bluuebunny/crossref_metadata_2025 huggingface dataset:
Radiative Properties of Numerically Generated Fractal Soot Aggregates: The Importance of Configuration Averaging
Authors: Liu Fengshan , Smallwood Gregory J.
Published: December 2009

The radiative properties of numerically generated fractal soot aggregates were studied using the numerically accurate generalized multisphere Mie-solution method. The fractal aggregates investigated in this study contain 10–600 primary particles of 30 nm in diameter. These fractal aggregates were numerically generated using a combination of the particle-cluster and cluster-cluster aggregation algorithms with fractal parameters representing flame-generated soot. Ten different realizations were obtained for a given aggregate size measured by the number of primary particles. The wavelength considered is 532 nm, and the corresponding size parameter of primary particle is 0.177. Attention is paid to the effect of different realizations of a fractal aggregate with identical fractal dimension, prefactor, primary particle diameter, and the number of primary particles on its orientation-averaged radiative properties. Most properties of practical interest exhibit relatively small variation with aggregate realization. However, other scattering properties, especially the vertical-horizontal differential scattering cross section, are very sensitive to the variation in geometrical configuration of primary particles. Orientation-averaged radiative properties of a single aggregate realization are not always sufficient to represent the properties of random-oriented ensemble of fractal aggregates.
Hamilton’s Principle of Entropy Production for Creep and Relaxation Processes
Authors: Yang Q. , Liu Y. R. , Bao J. Q.
Published: December 2009

In this paper, two subspaces of the state space of constrained equilibrium states for solids are proposed and addressed. One subspace, constrained affinity space, is conjugate-force space with fixed temperature and internal variable. It is revealed in this paper that the remarkable properties of the kinetic rate laws of scalar internal variables, established by Rice (1971, “Inelastic Constitutive Relations for Solids: An Internal Variable Theory and Its Application to Metal Plasticity,” J. Mech. Phys. Solids, 19, pp. 433–455) and elaborated by Yang (2005, “Normality Structures With Homogeneous Kinetic Rate Laws,” ASME J. Appl. Mech., 72, pp. 322–329; 2007, “Normality Structures With Thermodynamic Equilibrium Points,” ASME J. Appl. Mech., 74, pp. 965–971), are all located in constrained affinity space. Furthermore, the flow potential function monotonically increases along any ray from the origin in constrained affinity space. Another subspace, constrained configuration space, is the state space with fixed external variables. It is shown that the specific free and complementary energies monotonically decrease and increase, respectively, along the path of motion of the thermodynamic system of the material sample in constrained configuration space. For conservative conjugate forces, Hamilton’s action principle is established in constrained configuration space, and the action is the entropy production of the thermodynamic system in a time interval. The thermodynamic processes in constrained configuration space are just creep or relaxation processes of materials. The Hamilton principle can be considered as a fundamental principle of rheology.
Indirect Measurement of the Inertia Properties of a Knee Prosthesis Through a Simple Frequency-Domain Technique
Authors: Mucchi Emiliano , Bottoni Giuliamarta , Di Gregorio Raffaele
Published: December 2009

The dynamic study of humans carrying prostheses requires the rigid-body inertia properties of the prostheses. Since such properties are difficult to evaluate, in general, roughly estimated values of these quantities are used. These approximations may yield significant errors in the evaluation of some dynamic quantities (i.e., the inertia forces due to the prosthesis). This work is addressed to assess an experimental technique, based on frequency response function measurements, that indirectly measures the inertia properties of prostheses for transfemoral amputees. First, a specifically designed specimen and, then, a real prosthesis are tested for assessing the proposed technique. The results are that the measurement sensitivity is 0.002 kg m2 for inertia-tensor entries and 3 mm for center-of-gravity coordinates. Thus, the proposed technique is effective for a precise and fast evaluation of the inertia properties of medical devices such as prostheses.
A Haptic Simulator for Training the Application of Range of Motion Exercise to Premature Infants
Authors: Adnan Kareem N. , Ahmad Irfan , Coussens Maria , Eliakim Alon , Gallitto Susan , Grochow Donna , Koeppel Robin , Nemet Dan , Rich Julia , Waffarn Feizal , Cooper Dan M. , Reinkensmeyer David J.
Published: December 2009

The range of motion exercise is an experimental therapy for improving bone and muscle growth in premature infants but little is known about the magnitude of pressures that must be applied to the limbs during this exercise to elicit a physiological benefit and novice caregivers currently must rely on subjective instruction to learn to apply appropriate pressures. The goal of this study was to quantify the pressures applied by experienced caregivers during application of this exercise and to create a haptic simulator that could be used to train novice caregivers such as parents to apply the same pressures. We quantified the pressure applied by two neonatal intensive care nurses (“experts”) to the wrists of nine newborn, premature infants of varying gestational ages using an infant blood pressure cuff modified to act as a finger pressure sensor. The experts applied statistically significant different pressures depending on gestational age but did not differ significantly between themselves in the pressure they applied. We then created a robotic simulator of the premature infant wrist and programmed it to respond with the measured pressure-angle properties of the actual infants’ wrists. The novice adult participants (n=19) used the simulator to learn to apply target pressures for simulated wrists that corresponded to three different gestational ages. Training with the simulator for 30 min allowed the participants to learn to apply pressures significantly more like those of the experts. The performance improvement persisted at a retention test several days later. These results quantify for the first time the pressures applied during assisted exercise, include novel observations about joint flexibility and maturation early in life and suggest a strategy for teaching exercise intervention teams to provide assisted exercise within a more reproducible range using haptic simulation technology.
Design and Usability of a Home Telerehabilitation System to Train Hand Recovery Following Stroke
Authors: Durfee William K. , Weinstein Samantha A. , Bhatt Ela , Nagpal Ashima , Carey James R.
Published: December 2009

Current theories of stroke rehabilitation point toward paradigms of intense concentrated use of the afflicted limb as a means for motor program reorganization and partial function restoration. A home-based system for stroke rehabilitation that trains recovery of hand function by a treatment of concentrated movement was developed and tested. A wearable goniometer measured finger and wrist motions in both hands. An interface box transmitted sensor measurements in real-time to a laptop computer. Stroke patients used joint motion to control the screen cursor in a one-dimensional tracking task for several hours a day over the course of 10–14 days to complete a treatment of 1800 tracking trials. A telemonitoring component enabled a therapist to check in with the patient by video phone to monitor progress, to motivate the patient, and to upload tracking data to a central file server. The system was designed for use at home by patients with no computer skills. The system was placed in the homes of 20 subjects with chronic stroke and impaired finger motion, ranging from 2–305 mi away from the clinic, plus one that was a distance of 1057 miles. Fifteen subjects installed the system at home themselves after instruction in the clinic, while nine required a home visit to install. Three required follow-up visits to fix equipment. A post-treatment telephone survey was conducted to assess ease of use and most responded that the system was easy to use. Functional improvements were seen in the subjects enrolled in the formal treatment study, although the treatment period was too short to trigger cortical reorganization. We conclude that the system is feasible for home use and that tracking training has promise as a treatment paradigm.
Freebal: Design of a Dedicated Weight-Support System for Upper-Extremity Rehabilitation
Authors: Stienen Arno H. A. , Hekman Edsko E. G. , Prange Gerdienke B. , Jannink Michiel J. A. , van der Helm Frans C. T. , van der Kooij Herman
Published: December 2009

Most rehabilitation devices for the upper extremities include a weight-support system. In recent publications, weight support is shown to be effective for stroke rehabilitation. But current devices are often complex, have significant movement inertia, and/or limit the movement range. The goal of this study is to improve on current designs by introducing a novel, dedicated weight-support device, the Freebal. This passive mechanical device uses balanced spring mechanisms for constant-but-scalable forces to support the arm. It has a large workspace of roughly 1 m3, low movement impedance, and independent support at the elbow and wrist of up to 5 kg. An explorative cross-sectional study with eight patients shows the Freebal to instantly extend the range of motion of the affected arm by 7%. In conclusion, most requirements are met for patients to benefit from therapy with the Freebal, potentially progressing earlier to more motivating, functional training.
Delivery of MicroRNA-126 by Apoptotic Bodies Induces CXCL12-Dependent Vascular Protection
Authors: Zernecke Alma , Bidzhekov Kiril , Noels Heidi , Shagdarsuren Erdenechimeg , Gan Lin , Denecke Bernd , Hristov Mihail , Köppel Thomas , Jahantigh Maliheh Nazari , Lutgens Esther , Wang Shusheng , Olson Eric N. , Schober Andreas , Weber Christian
Published: December 2009

Apoptotic endothelial cells release microRNA-containing microvesicles to modulate the responses of neighboring cells and reduce atherosclerosis in mice.
Only a Subset of Met-Activated Pathways Are Required to Sustain Oncogene Addiction
Authors: Bertotti Andrea , Burbridge Mike F. , Gastaldi Stefania , Galimi Francesco , Torti Davide , Medico Enzo , Giordano Silvia , Corso Simona , Rolland-Valognes Gaëlle , Lockhart Brian P. , Hickman John A. , Comoglio Paolo M. , Trusolino Livio
Published: December 2009

Cells addicted to different oncogenic receptor tyrosine kinases develop common downstream mechanisms to sustain malignancy.
Traversing the Valley of Death: A Guide to Assessing Prospects for Translational Success
Authors: Coller Barry S. , Califf Robert M.
Published: December 2009

In an attempt to help investigators judge the likelihood that a translational project will be successful, we propose a series of questions that form a self-guided global risk assessment tool.
Foot orthoses and gait: a systematic review and meta-analysis of literature pertaining to potential mechanisms
Authors: Mills Kathryn , Blanch Peter , Chapman Andrew R , McPoil Thomas G , Vicenzino Bill
Published: December 2009

This article systematically reviews the available literature to improve our understanding of the physiological basis for orthoses under the kinematic, shock attenuation and neuromotor control paradigms. The propositions made under these three paradigms have not been systematically reviewed collectively, and as such, there is no single-point synthesis of this clinically relevant body of evidence and somewhat disparate findings. Our comprehensive search strategy yielded 22 papers. Under each paradigm, the role of orthoses with different design features including combinations of posting, moulding and density was analysed. Where possible, data have been pooled to provide an increased level of confidence in findings. The main findings in the kinematic paradigm were that posted non-moulded orthoses systematically reduced peak rearfoot eversion (2.12° (95% CI 0.72 to 3.53)) and tibial internal rotation (1.33° (0.12 to 2.53)) in non-injured cohorts. In the shock attenuation paradigm, it was found that non-posted moulded and posted moulded orthoses produced large reductions in loading rate and vertical impact force when compared with a control and to a posted non-moulded orthosis. The neuromotor control paradigm seems to be the least conclusive in its outcome. Based on our review, this paper concludes with rudimentary guidelines for the prescription of orthosis, that sports medicine practitioners may use in their clinical decision-making process. The need for further research focusing on the role of injury, particularly in neuromotor control modification and long-term adaptation to orthoses, was highlighted.
Metabolic functions of atypical protein kinase C: “good” and “bad” as defined by nutritional status
Authors: Farese Robert V. , Sajan Mini P.
Published: December 2009

Atypical protein kinase C (aPKC) isoforms mediate insulin effects on glucose transport in muscle and adipose tissues and lipid synthesis in liver and support other metabolic processes, expression of enzymes needed for islet insulin secretion and hepatic glucose production/release, CNS appetite suppression, and inflammatory responses. In muscle, selective aPKC deficiency impairs glucose uptake and produces insulin resistance and hyperinsulinemia, which, by activating hepatic aPKC, provokes inordinate increases in lipid synthesis and produces typical “metabolic syndrome” features. In contrast, hepatic aPKC deficiency diminishes lipid synthesis and protects against metabolic syndrome features. Unfortunately, aPKC is deficient in muscle but paradoxically conserved in liver in obesity and type 2 diabetes mellitus; this combination is particularly problematic because it promotes lipid and carbohydrate abnormalities. Accordingly, metabolic effects of aPKCs can be “good” or “bad,” depending upon nutritional status; thus, muscle glucose uptake, islet insulin secretion, hepatic glucose and lipid production/release, and adipose fat synthesis/storage would be important for survival during periods of limited food availability and therefore be “good.” However, during times of food surfeit, excessive activation of hepatic aPKC, whether caused by overnutrition or impairments in extrahepatic effects of insulin, would lead to inordinate increases in hepatic lipid synthesis and metabolic syndrome features and therefore be “bad.” In keeping with these ideas, the inhibition of hepatic aPKC markedly ameliorates lipid and carbohydrate abnormalities in experimental models of obesity and type 2 diabetes. We postulate that a similar approach may be useful for treating humans.
On the role of glucose-dependent insulintropic polypeptide in postprandial metabolism in humans
Authors: Asmar Meena , Tangaa Winnie , Madsbad Sten , Hare Kristine , Astrup Arne , Flint Anne , Bülow Jens , Holst Jens Juul
Published: December 2009

We investigated the role of glucose-dependent insulintropic polypeptide (GIP) in the regulation of gastric emptying (GE), appetite, energy intake (EI), energy expenditure (EE), plasma levels of triglycerides (TAG), and free fatty acids (FFA) in humans. First, 20 healthy males received intravenous infusion of GIP (0.8 pmol·kg−1·min−1) or saline for 300 min during and after a fixed meal ( protocol 1). GE was measured using paracetamol, appetite sensations using visual analog scales, EE using indirect calorimetry, and EI during a subsequent ad libitum meal (at 300 min). Next, 10 healthy males received intravenous infusions of Intralipid, glucose, or Intralipid plus glucose, with and without GIP (1.5 pmol·kg−1·min−1) for 300 min ( protocol 2). In protocol 1, GIP did not have any effect on GE, EI, EE, removal of TAG, or FFA and did not influence the subjective feeling of hunger, satiety, fullness or prospective food consumption compared with saline. In protocol 2, no difference was seen in the plasma TAG on Intralipid + GIP/saline and Intralipid + glucose + GIP/saline days. FFA concentrations were lower on Intralipid + glucose + GIP/saline days ( P < 0.05) compared with Intralipid + GIP/saline days and on Intralipid + GIP day ( P < 0.004) compared with Intralipid + saline day. Insulin increased on all GIP days compared with saline days ( P < 0.05). In conclusion, while confirming its insulinotropic effects, these data suggest that GIP does not affect GE, appetite, energy intake, EE, or the clearance rate of the applied TAG formulation in humans. However, both insulin and GIP lower post-Intralipid FFA concentration, GIP probably via stimulation of insulin secretion, increasing FFA reesterification.
PROC QTL—A SAS Procedure for Mapping Quantitative Trait Loci
Authors: Hu Zhiqiu , Xu Shizhong
Published: December 2009

Statistical analysis system (SAS) is the most comprehensive statistical analysis software package in the world. It offers data analysis for almost all experiments under various statistical models. Each analysis is performed using a particular subroutine, called a procedure (PROC). For example, PROC ANOVA performs analysis of variances. PROC QTL is a user‐defined SAS procedure for mapping quantitative trait loci (QTL). It allows users to perform QTL mapping for continuous and discrete traits within the SAS platform. Users of PROC QTL are able to take advantage of all existing features offered by the general SAS software, for example, data management and graphical treatment. The current version of PROC QTL can perform QTL mapping for all line crossing experiments using maximum likelihood (ML), least square (LS), iteratively reweighted least square (IRLS), Fisher scoring (FISHER), Bayesian (BAYES), and empirical Bayes (EBAYES) methods.
Repeated Shadow Track Orbits for Space‐SunSetter Missions
Authors: Gad Ahmed , Abdelkhalik Ossama
Published: December 2009

This paper introduces a new set of orbits, the “Repeated Shadow Track Orbits.” In these orbits, the shadow of a spacecraft on the Earth visits the same locations periodically every desired number of days. TheJ2perturbation is utilized to synchronize the spacecraft shadow motion with both the Earth rotational motion and the Earth‐Sun vector rotation. Motivation for the design of new shadow track orbits comes from the need to save energy. The general mathematical model to design a Repeated Shadow Track Orbit (RSTO) is presented within this paper. RSTOs′ conditions are formulated and numerically solved. Results show the feasibility of RSTOs. An optimization process is also developed to maximize the shadow duration over a given site. A Genetic Algorithm (GA) technique is utilized for optimization.
Detection of GAD65 Autoreactive T‐Cells by HLA Class I Tetramers in Type 1 Diabetic Patients
Authors: Giuliani Laura , Mele Raffaella , Di Giovine Monica , Altieri Laura , Crinò Antonino , Ravà Lucilla , Fierabracci Alessandra
Published: December 2009

Type 1 diabetes (T1D) is an autoimmune disease, in which pancreatic β cells are destroyed in genetically predisposed individuals. While the direct contribution of autoantibodies to the disease pathogenesis is controversial, it is generally recognised that the mechanism of β cell destruction is mediated by autoreactive T cells that had escaped the thymic selection. We aimed to design a method to detect circulating CD8+ T cells autoreactive against an epitope of the glutamic acid decarboxylase autoantigen, isoform 65 (GAD65) ex vivo in T1D patients by using HLA class I tetramers. Low frequencies of GAD65 peptide‐specific CD8+ cytotoxic T lymphocytes were detected in peripheral blood lymphocytes (PBMC) of normal controls after GAD65 peptide‐specific stimulation. Conversely, their frequencies were significantly higher than in controls in PBMC of T1D patients after GAD65 peptide stimulation. These preliminary data are encouraging in order to develop a reliable assay to be employed in large‐scale screening studies.
Immune Stimulatory Receptor CD40 Is Required for T-Cell Suppression and T Regulatory Cell Activation Mediated by Myeloid-Derived Suppressor Cells in Cancer
Authors: Pan Ping-Ying , Ma Ge , Weber Kaare J. , Ozao-Choy Junko , Wang George , Yin Bingjiao , Divino Celia M. , Chen Shu-Hsia
Published: December 2009

Immune tolerance to tumors is often associated with accumulation of myeloid-derived suppressor cells (MDSC) and an increase in the number of T-regulatory cells (Treg). In tumor-bearing mice, MDSCs can themselves facilitate the generation of tumor-specific Tregs. In this study, we demonstrate that expression of the immune stimulatory receptor CD40 on MDSCs is required to induce T-cell tolerance and Treg accumulation. In an immune reconstitution model, adoptive transfer of Gr-1+CD115+ monocytic MDSCs derived from CD40-deficient mice failed to recapitulate the ability of wild-type MDSCs to induce tolerance and Treg development in vivo. Agonistic anti-CD40 antibodies phenocopied the effect of CD40 deficiency and also improved the therapeutic efficacy of IL-12 and 4-1BB immunotherapy in the treatment of advanced tumors. Our findings suggest that CD40 is essential not only for MDSC-mediated immune suppression but also for tumor-specific Treg expansion. Blockade of CD40-CD40L interaction between MDSC and Treg may provide a new strategy to ablate tumoral immune suppression and thereby heighten responses to immunotherapy. Cancer Res; 70(1); 99–108
Chemosensitivity Profiles Identify Polymorphisms in the p53 Network Genes 14-3-3τ and CD44 That Affect Sarcoma Incidence and Survival
Authors: Vazquez Alexei , Grochola Lukasz F. , Bond Elisabeth E. , Levine Arnold J. , Taubert Helge , Müller Thomas H. , Würl Peter , Bond Gareth L.
Published: December 2009

The p53 regulatory network responds to cellular stresses by initiating processes such as cell cycle arrest and apoptosis. These responses inhibit cellular transformation and mediate the response to many forms of cancer therapies. Functional variants in the genes comprising this network could help identify individuals at greater risk for cancer and patients with poorer responses to therapies, but few such variants have been identified as yet. We use the NCI60 human tumor cell line anticancer drug screen in a scan of single nucleotide polymorphisms (SNP) in 142 p53 stress response genes and identify 7 SNPs that exhibit allelic differences in cellular responses to a large panel of cytotoxic chemotherapeutic agents. The greatest differences are observed for SNPs in 14-3-3τ (YWHAQ; rs6734469, P = 5.6 × 10−47) and CD44 (rs187115, P = 8.1 × 10−24). In soft-tissue sarcoma patients, we find that the alleles of these SNPs that associate with weaker growth responses to chemotherapeutics associate with poorer overall survival (up to 2.89 relative risk, P = 0.011) and an earlier age of diagnosis (up to 10.7 years earlier, P = 0.002). Our findings define genetic markers in 14-3-3τ and CD44 that might improve the treatment and prognosis of soft-tissue sarcomas. Cancer Res; 70(1); 172–80
Adult-Type Granulosa Cell Tumors and FOXL2 Mutation
Authors: Köbel Martin , Gilks C. Blake , Huntsman David G.
Published: December 2009

Little is known about the pathogenesis of ovarian granulosa cell tumors. Recently, we reported the identification of a somatic FOXL2 402C→G mutation that is present in virtually all adult-type granulosa cell tumors, but not in a wide range of other tumor types. This finding has important implications for the diagnosis and classification of ovarian sex cord-stromal tumors, provides insight into the pathogenesis of adult-type granulosa cell tumors, and opens possibilities for the development of targeted therapies. [Cancer Res 2009;69(24):9160–2]
A Proteomic Approach for the Identification of Vascular Markers of Liver Metastasis
Authors: Borgia Beatrice , Roesli Christoph , Fugmann Tim , Schliemann Christoph , Cesca Marta , Neri Dario , Giavazzi Raffaella
Published: December 2009

Vascular proteins expressed at liver metastasis sites could serve as prognostic markers or as targets for pharmacodelivery applications. We employed a proteomic approach to define such proteins in three syngeneic mouse models of liver metastasis. Vascular structures were biotinylated in vivo by a terminal perfusion technique, followed by mass spectrometric analysis of accessible biotinylated proteins. In this manner, we identified 12 proteins for which expression was selectively associated with liver metastasis, confirming this association by tissue immunofluorescence or in vivo localization with radiolabeled antibodies. In summary, our findings identify vascular proteins that may have prognostic or drug-targeting use in addressing liver metastases, a common issue in many advanced cancers. Cancer Res; 70(1); 309–18
Increased Prevalence of Precursor Lesions in Familial Pancreatic Cancer Patients
Authors: Shi Chanjuan , Klein Alison P. , Goggins Michael , Maitra Anirban , Canto Marcia , Ali Syed , Schulick Richard , Palmisano Emily , Hruban Ralph H.
Published: December 2009

Purpose: Histologic findings in 51 pancreata resected from patients with a strong family history of pancreatic cancer were compared with the findings in 40 pancreata resected from patients with sporadic pancreatic cancer. None of the patients in the familial group had a known inherited syndrome other than familial pancreatic cancer.

Experimental Design: Precursor lesions, including pancreatic intraepithelial neoplasia (PanIN), intraductal papillary mucinous neoplasm (IPMN), and incipient IPMN, were quantified. Invasive cancers were classified using established histologic criteria.

Results: The individual precursor lesions identified in both groups were histologically similar. Precursor lesions were more common in the familial cases than in the sporadic cases. The relative rate of PanINs per square centimeter was 2.75-fold higher (95% confidence interval, 2.05-3.70; adjusted for age) in familial compared with sporadic cases. PanIN-3 lesions were more common in familial versus sporadic pancreatic cancer patients (relative rate, 4.20; 95% confidence interval, 2.22-7.93; adjusted for age). High-grade incipient IPMNs were only observed in the familial cases. Nine of the 51 (18%) familial pancreatic cancers and 4 of the 40 (10%) sporadic cancers arose in association with an IPMN. No significant differences were found in the types of invasive cancers.

Conclusions: Noninvasive precursor lesions are more common in patients with a strong family history of pancreatic cancer than in patients with sporadic disease, and precursor lesions are of a higher grade in patients with a strong family history of pancreatic cancer. These findings can form a basis for the design of screening tests for the early detection of pancreatic neoplasia. (Clin Cancer Res 2009;15(24):7737–43)
Transforming Growth Factor-β–Stimulated Clone-22 Is an Androgen-Regulated Gene That Enhances Apoptosis in Prostate Cancer following Insulin-Like Growth Factor-I Receptor Inhibition
Authors: Sprenger Cynthia C.T. , Haugk Kathleen , Sun Shihua , Coleman Ilsa , Nelson Peter S. , Vessella Robert L. , Ludwig Dale L. , Wu Jennifer D. , Plymate Stephen R.
Published: December 2009

Purpose: Inhibition of insulin-like growth factor (IGF) signaling using the human IGF-I receptor monoclonal antibody A12 is most effective at inducing apoptosis in prostate cancer xenografts in the presence of androgen. We undertook this study to determine mechanisms for increased apoptosis by A12 in the presence of androgens.

Experimental Methods: The castrate-resistant human xenograft LuCaP 35 V was implanted into intact or castrate severe combined immunodeficient mice and treated with A12 weekly. After 6 weeks of tumor growth, animals were sacrificed and tumors were removed and analyzed for cell cycle distribution/apoptosis and cDNA arrays were done.

Results: In castrate mice, the tumors were delayed in G2 with no apoptosis; in contrast, tumors from intact mice underwent apoptosis with either G1 or G2 delay. Transforming growth factor-β–stimulated clone-22 (TSC-22) was significantly elevated in tumors from the intact mice compared with castrate mice, especially in those tumors with the highest levels of apoptosis. To further determine the function of TSC-22, we transfected various human prostate cancer cell lines with a plasmid expressing TSC-22. Cell lines overexpressing TSC-22 showed an increase in apoptosis and a delay in G1. When these cell lines were placed subcutaneously in athymic nude mice, a decreased number of animals formed tumors and the rate of tumor growth was decreased compared with control tumors.

Conclusions: These data indicate that IGF-I receptor inhibition in the presence of androgen has an enhanced effect on decreasing tumor growth, in part, through increased expression of the tumor suppressor gene TSC-22. (Clin Cancer Res 2009;15(24):7634–41)
Noninvasive Prediction of Fracture Risk in Patients with Metastatic Cancer to the Spine
Authors: Snyder Brian D. , Cordio Marsha A. , Nazarian Ara , Kwak S. Daniel , Chang David J. , Entezari Vahid , Zurakowski David , Parker Leroy M.
Published: December 2009

Purpose: Skeletal metastases affect up to 85% of breast cancer patients by the time of their death. This prospective in vivo study evaluated the diagnostic performance of computed tomography–based structural rigidity analysis (CTRA) to predict vertebral fracture risk in breast cancer patients with skeletal metastasis and in comparison with the current standard of care.

Experimental Design: Torso CT scans of 94 women with vertebral metastatic breast cancer were obtained as part of routine screening for lung and liver metastases. The load-bearing capacity (LBC) and axial (EA) and bending (EI) rigidities of vertebrae T8 to L5 were calculated from CT images. The LBC was normalized by patient body mass index (BMI) to account for height and mass variations. Vertebral fracture risk was also calculated using the current radiographic-based criteria based on lesion size and location. The actual occurrence of a new vertebral fracture was assessed radiographically over the ensuing 4 months.

Results: Eleven vertebral fractures occurred in 10 patients. The structural parameters EA, EI, LBC, and LBC/BMI were all 100% sensitive and 55%, 53%, 44%, and 70% specific to predict fracture risk, respectively. Although radiographic criteria correctly predicted all fracture cases (100% sensitive), only 48 of the 236 spinal segments that did not have a fracture were correctly predicted not to fracture (20% specific).

Conclusions: CTRA, using CT scans as part of routine screening for lung and liver metastasis, is shown to be as sensitive as, and significantly more specific than, the current radiographic criteria for predicting vertebral fracture in breast cancer patients with skeletal metastasis. (Clin Cancer Res 2009;15(24):7676–83)
The Ligamp TP53 Assay for Detection of Minimal Residual Disease in Head and Neck Squamous Cell Carcinoma Surgical Margins
Authors: Poeta M. Luana , Manola Judith , Goldenberg David , Forastiere Arlene , Califano Joseph A. , Ridge John A. , Goodwin Jarrard , Kenady Daniel , Saunders John , Westra William , Sidransky David , Koch Wayne M.
Published: December 2009

Purpose: Detect tumor-related DNA using LigAmp in histologically clear margins and associate results with clinical outcome.

Experimental Design: Patients with head and neck cancer were registered for molecular analysis of surgical margins. Adequacy of resection was ensured using histologic margin analysis. Further margins were then harvested and DNA extracted. TP53 mutations in tumor were determined using Affymetrix p53 GeneChip. Margins were analyzed by Ligamp in comparison with standard curves for quantification of mutant DNA. Ligation used two oligonucleotides to isolate DNA targeting the mutation. Ligated DNA was amplified using real-time PCR. The quantity of mutation in the margin was determined as percent of mutant species relative to plasmid and relative to tumor. Cutpoints were identified and defined groups were evaluated for local failure-free, cancer-specific, and overall survival. Study margins were examined for presence of tumor by light microscopy.

Results: Tissue from 95 patients with common mutations was analyzed. Fifteen experienced local recurrence. Cutpoints of 0.15% for mutant species relative to plasmid and 0.5% for mutant species relative to tumor were chosen as most selective of recurrent cases. LigAmp had slightly better area under the receiver operator characteristic curve (P = 0.09) than light microscopy correctly predicting 9 of 15 recurrent tumors. There were 6 false negative cases and 26 false positive results. No statistically significant distinctions were observed in cancer-specific or overall survival in this limited cohort.

Conclusions: Ligamp provides quantifiable, sensitive detection of mutant DNA in histologically normal margins. Detection of mutant species in margins may identify patients at risk of local recurrence. (Clin Cancer Res 2009;15(24):7658–65)
Relationship between Topoisomerase 2A RNA Expression and Recurrence after Adjuvant Chemotherapy for Breast Cancer
Authors: Sparano Joseph A. , Goldstein Lori J. , Childs Barrett H. , Shak Steven , Brassard Diana , Badve Sunil , Baehner Frederick L. , Bugarini Roberto , Rowley Steve , Perez Edith , Shulman Lawrence N. , Martino Silvana , Davidson Nancy E. , Sledge George W. , Gray Robert
Published: December 2009

Purpose: To perform an exploratory analysis of the relationship between gene expression and recurrence in operable hormone receptor (HR)–positive, human epidermal growth factor receptor 2 (HER2)–normal breast cancer patients treated with adjuvant doxorubicin-containing chemotherapy.

Experimental Design: RNA was extracted from archived tumor samples derived from 378 patients with stage I to III HR-positive, HER2-normal breast cancer and analyzed by reverse transcription-PCR for a panel of 374 genes, including the 21-gene recurrence score (RS). Patients were randomized to receive adjuvant doxorubicin plus cyclophosphamide or docetaxel in trial E2197, with no difference in recurrence seen in the treatment arms. All available recurrent cases were selected plus a nonrecurrent cohort. Cox proportional hazard models were used to identify relationships between gene expression and recurrence.

Results: TOP2A expression exhibited the strongest association with increased recurrence risk (P = 0.01), and was significantly associated with recurrence (P = 0.008) in a multivariate analysis adjusted for clinicopathologic features. Elevated TOP2A expression above the median was associated with a 2.6-fold increase (95% confidence interval, 1.3-5.2; P = 0.008) in risk of recurrence if the RS was <18, and a 2.0-fold increase (95% confidence interval, 1.2-3.2, P = 0.003) if there was an intermediate RS of 18 to 30.

Conclusions: In patients with HR-positive, HER2-normal breast cancer, a population known to have a low incidence of TOP2A gene alterations thought to be predictive of anthracycline benefit, there is a range of TOP2A RNA expression that is strongly associated with recurrence after adjuvant anthracyclines, which provides information complementary to RS, indicating that it merits further evaluation as a prognostic and predictive marker. (Clin Cancer Res 2009;15(24):7693–700)
Clinical Benefit of Allogeneic Melanoma Cell Lysate–Pulsed Autologous Dendritic Cell Vaccine in MAGE-Positive Colorectal Cancer Patients
Authors: Toh Han Chong , Wang Who-Whong , Chia Whay Kuang , Kvistborg Pia , Sun Li , Teo Kelly , Phoon Yee Peng , Soe Yatanar , Tan Sze Huey , Hee Siew Wan , Foo Kian Fong , Ong Simon , Koo Wen Hsin , Zocca Mai-Britt , Claesson Mogens H.
Published: December 2009

Purpose: We evaluated the clinical benefit of an allogeneic melanoma cell lysate (MCL)–pulsed autologous dendritic cell (DC) vaccine in advanced colorectal cancer patients expressing at least one of six MAGE-A antigens overexpressed by the cell line source of the lysate.

Experimental Design: DCs were cultured from peripheral blood mononuclear cells (PBMC), pulsed with the allogeneic MCL, and matured using cytokines that achieved high CD83- and CCR7-expressing DCs. Each patient received up to 10 intradermal vaccinations (3-5 × 106 cells per dose) at biweekly intervals.

Results: Twenty patients received a total of 161 vaccinations. Treatment was well tolerated and quality of life measurements did not vary much across time. One patient experienced partial response [5%; 95% confidence interval (CI), 1-24%] and seven achieved stable disease (35%; 95% CI, 18-57%), one of whom also achieved late tumor regression, yielding a clinical benefit response rate of 40% (95% CI, 22-61%). Although overall median progression-free survival was 2.4 months (95% CI, 1.9-4.1 months), five patients (25%) experienced prolonged progression-free survival (>6 months), two of whom (10%) remain progression-free for >27 and >37 months, respectively. This result is particularly meaningful as all patients had progressive disease before treatment. Overall, DC vaccination was associated with a serial decline in regulatory T cells. Using an antibody array, we characterized plasma protein profiles in responding patients that may correlate with vaccine activity and report a prevaccination protein signature distinguishing responders from nonresponders.

Conclusion: This phase II vaccine study using mature, MCL-pulsed DCs has shown promising results and warrants further evaluation in a prospective randomized setting. (Clin Cancer Res 2009;15(24):7726–36)
Phase II Trial of Temozolomide and Sorafenib in Advanced Melanoma Patients with or without Brain Metastases
Authors: Amaravadi Ravi K. , Schuchter Lynn M. , McDermott David F. , Kramer Amy , Giles Lydia , Gramlich Kristi , Carberry Mary , Troxel Andrea B. , Letrero Richard , Nathanson Katherine L. , Atkins Michael B. , O'Dwyer Peter J. , Flaherty Keith T.
Published: December 2009

Purpose: The combination of the oral alkylating agent temozolomide and the oral multikinase inhibitor sorafenib was evaluated in advanced melanoma patients.

Experimental Design: Patients with metastatic melanoma (n = 167) were treated on four arms. All patients received sorafenib at 400 mg p.o. twice daily without interruption. Patients without brain metastases or prior temozolomide were randomized between arm A: extended dosing of temozolomide (75 mg/m2 temozolomide daily for 6 of every 8 weeks) and arm B: standard dosing (150 mg/m2 temozolomide daily for 5 of every 28 days). Patients previously treated with temozolomide were enrolled on arm C: extended dosing of temozolomide. Patients with brain metastases and no prior temozolomide were assigned to arm D: standard dosing. The primary end point was 6-month progression-free survival (PFS) rate. Secondary end points included response rate, toxicity rates, and the rates of BRAF or NRAS mutations.

Results: The 6-month PFS rate for arms A, B, C, and D were 50%, 40%, 11%, and 23%. The median PFS for patients on arm A, B, C, and D was 5.9, 4.2, 2.2, and 3.5 months, respectively. No significant differences were observed between arms A and B in 6-month PFS rate, median PFS, or response rates. Treatment was well tolerated in all arms. No significant differences in toxicity were observed between arms A and B except for more grade 3 to 4 lymphopenia in arm A.

Conclusion: Temozolomide plus sorafenib was well tolerated and showed activity in melanoma patients without prior history of temozolomide. The activity of this combination regimen warrants further investigation. (Clin Cancer Res 2009;15(24):7711–8)
In vivo activation of the hypoxia-targeted cytotoxin AQ4N in human tumor xenografts
Authors: Williams Kaye J. , Albertella Mark R. , Fitzpatrick Brian , Loadman Paul M. , Shnyder Steven D. , Chinje Edwin C. , Telfer Brian A. , Dunk Chris R. , Harris Peter A. , Stratford Ian J.
Published: December 2009

AQ4N (banoxantrone) is a prodrug that, under hypoxic conditions, is enzymatically converted to a cytotoxic DNA-binding agent, AQ4. Incorporation of AQ4N into conventional chemoradiation protocols therefore targets both oxygenated and hypoxic regions of tumors, and potentially will increase the effectiveness of therapy. This current pharmacodynamic and efficacy study was designed to quantify tumor exposure to AQ4 following treatment with AQ4N, and to relate exposure to outcome of treatment. A single dose of 60 mg/kg AQ4N enhanced the response of RT112 (bladder) and Calu-6 (lung) xenografts to treatment with cisplatin and radiation therapy. AQ4N was also given to separate cohorts of tumor-bearing mice 24 hours before tumor excision for subsequent analysis of metabolite levels. AQ4 was detected by high performance liquid chromatography/mass spectrometry in all treated samples of RT112 and Calu-6 tumors at mean concentrations of 0.23 and 1.07 μg/g, respectively. These concentrations are comparable with those shown to be cytotoxic in vitro. AQ4-related nuclear fluorescence was observed in all treated tumors by confocal microscopy, which correlated with the high performance liquid chromatography/mass spectrometry data. The presence of the hypoxic marker Glut-1 was shown by immunohistochemistry in both Calu-6 tumors and RT112 tumors, and colocalization of AQ4 fluorescence and Glut-1 staining strongly suggested that AQ4N was activated in these putatively hypoxic areas. This is the first demonstration that AQ4N will increase the efficacy of chemoradiotherapy in preclinical models; the intratumoral levels of AQ4 found in this study are comparable with tumor AQ4 levels found in a recent phase I clinical study, which suggests that these levels could be potentially therapeutic. [Mol Cancer Ther 2009;8(12):3266–75]
BMS-754807, a small molecule inhibitor of insulin-like growth factor-1R/IR
Authors: Carboni Joan M. , Wittman Mark , Yang Zheng , Lee Francis , Greer Ann , Hurlburt Warren , Hillerman Stephen , Cao Carolyn , Cantor Glenn H. , Dell-John Janet , Chen Cliff , Discenza Lorell , Menard Krista , Li Aixin , Trainor George , Vyas Dolatrai , Kramer Robert , Attar Ricardo M. , Gottardis Marco M.
Published: December 2009

BMS-754807 is a potent and reversible inhibitor of the insulin-like growth factor 1 receptor/insulin receptor family kinases (Ki, <2 nmol/L). It is currently in phase I development for the treatment of a variety of human cancers. BMS-754807 effectively inhibits the growth of a broad range of human tumor types in vitro, including mesenchymal (Ewing's, rhabdomyosarcoma, neuroblastoma, and liposarcoma), epithelial (breast, lung, pancreatic, colon, gastric), and hematopoietic (multiple myeloma and leukemia) tumor cell lines (IC50, 5–365 nmol/L); the compound caused apoptosis in a human rhabdomyosarcoma cell line, Rh41, as shown by an accumulation of the sub-G1 fraction, as well as by an increase in poly ADP ribose polymerase and Caspase 3 cleavage. BMS-754807 is active in vivo in multiple (epithelial, mesenchymal, and hematopoietic) xenograft tumor models with tumor growth inhibition ranging from 53% to 115% and at a minimum effective dose of as low as 6.25 mg/kg dosed orally daily. Combination studies with BMS-754807 have been done on multiple human tumor cell types and showed in vitro synergies (combination index, <1.0) when combined with cytotoxic, hormonal, and targeted agents. The combination of cetuximab and BMS-754807 in vivo, at multiple dose levels, resulted in improved clinical outcome over single agent treatment. These data show that BMS-754807 is an efficacious, orally active growth factor 1 receptor/insulin receptor family–targeted kinase inhibitor that may act in combination with a wide array of established anticancer agents. [Mol Cancer Ther 2009;8(12):3341–9]
Sulindac sulfide selectively inhibits growth and induces apoptosis of human breast tumor cells by phosphodiesterase 5 inhibition, elevation of cyclic GMP, and activation of protein kinase G
Authors: Tinsley Heather N. , Gary Bernard D. , Keeton Adam B. , Zhang Wei , Abadi Ashraf H. , Reynolds Robert C. , Piazza Gary A.
Published: December 2009

Sulindac displays promising antineoplastic activity, but toxicities from cyclooxygenase inhibition limit its use for chemoprevention. Previous reports suggest that its anticancer properties may be attributed to a cyclooxygenase-independent mechanism, although alternative targets have not been well defined. Here, we show that sulindac sulfide (SS) induces apoptosis and inhibits the growth of human breast tumor cells with IC50 values of 60 to 85 μmol/L. Within the same concentration range, SS inhibited cyclic GMP (cGMP) hydrolysis in tumor cell lysates but did not affect cyclic AMP hydrolysis. SS did not induce apoptosis of normal human mammary epithelial cells (HMEC) nor did it inhibit phosphodiesterase (PDE) activity in HMEC lysates. SS increased intracellular cGMP levels and activated protein kinase G in breast tumor cells but not HMEC. The guanylyl cyclase (GC) activator, NOR-3, and cGMP PDE inhibitors, trequinsin and MY5445, displayed similar growth-inhibitory activity as SS, but the adenylyl cyclase activator, forskolin, and other PDE inhibitors had no effect. Moreover, GC activation increased the sensitivity of tumor cells to SS, whereas GC inhibition reduced sensitivity. By comparing PDE isozyme profiles in breast tumor cells with HMEC and determining the sensitivity of recombinant PDE isozymes to SS, PDE5 was found to be overexpressed in breast tumor cells and selectively inhibited by SS. The mechanism of SS binding to the catalytic domain of PDE5 was revealed by molecular modeling. These data suggest that PDE5 inhibition is responsible for the breast tumor cell growth–inhibitory and apoptosis-inducing activity of SS and may contribute to the chemopreventive properties of sulindac. [Mol Cancer Ther 2009;8(12):3331–40]
RanBPM Has Proapoptotic Activities That Regulate Cell Death Pathways in Response to DNA Damage
Authors: Atabakhsh Elnaz , Bryce Dawn M. , Lefebvre Karen J. , Schild-Poulter Caroline
Published: December 2009

Ran-binding protein M (RanBPM) is a nucleocytoplasmic protein previously implicated in various signaling pathways, but whose function remains enigmatic. Here, we provide evidence that RanBPM functions as an activator of apoptotic pathways induced by DNA damage. First, transient expression of RanBPM in HeLa cells induced cell death through caspase activation, and in the long-term, forced expression of RanBPM impaired cell viability. RanBPM COOH-terminal domain stimulated the ability of RanBPM to induce caspase activation, whereas this activity was negatively regulated by the central SPRY domain. Second, small interfering RNA–directed knockdown of RanBPM prevented DNA damage–induced apoptosis, as evidenced by the marked reduction in caspase-3 and caspase-2 activation. This correlated with a magnitude fold increase in the survival of RanBPM-depleted cells. Following ionizing radiation treatment, we observed a progressive relocalization of RanBPM from the nucleus to the cytoplasm, suggesting that the activation of apoptotic pathways by RanBPM in response to ionizing radiation may be regulated by nucleocytoplasmic trafficking. Finally, RanBPM downregulation was associated with a marked decrease of mitochondria-associated Bax, whereas Bcl-2 overall levels were dramatically upregulated. Overall, our results reveal a novel proapoptotic function for RanBPM in DNA damage–induced apoptosis through the regulation of factors involved in the mitochondrial apoptotic pathway. (Mol Cancer Res 2009;7(12):1962–72)