bluuebunny/crossref_metadata_2025 huggingface dataset:
The influence of a 5-wk whole body vibration on electrophysiological properties of rat hindlimb spinal motoneurons
Authors: Bączyk M. , Hałuszka A. , Mrówczyński W. , Celichowski J. , Krutki P.
Published: March 2013

The study aimed at determining the influence of a whole body vibration (WBV) on electrophysiological properties of spinal motoneurons. The WBV training was performed on adult male Wistar rats, 5 days a week, for 5 wk, and each daily session consisted of four 30-s runs of vibration at 50 Hz. Motoneuron properties were investigated intracellularly during experiments on deeply anesthetized animals. The experimental group subjected to the WBV consisted of seven rats, and the control group of nine rats. The WBV treatment induced no significant changes in the passive membrane properties of motoneurons. However, the WBV-evoked adaptations in excitability and firing properties were observed, and they were limited to fast-type motoneurons. A significant decrease in rheobase current and a decrease in the minimum and the maximum currents required to evoke steady-state firing in motoneurons were revealed. These changes resulted in a leftward shift of the frequency-current relationship, combined with an increase in slope of this curve. The functional relevance of the described adaptive changes is the ability of fast motoneurons of rats subjected to the WBV to produce series of action potentials at higher frequencies in a response to the same intensity of activation. Previous studies proved that WBV induces changes in the contractile parameters predominantly of fast motor units (MUs). The data obtained in our experiment shed a new light to possible explanation of these results, suggesting that neuronal factors also play a substantial role in MU adaptation.
Strain-specific variations in cation content and transport in mouse erythrocytes
Authors: Rivera Alicia , Zee Robert Y. L. , Alper Seth L. , Peters Luanne L. , Brugnara Carlo
Published: March 2013

Studies of ion transport pathophysiology in hematological disorders and tests of possible new therapeutic agents for these disorders have been carried out in various mouse models because of close functional similarities between mouse and human red cells. We have explored strain-specific differences in erythrocyte membrane physiology in 10 inbred mouse strains by determining erythrocyte contents of Na+, K+, and Mg2+, and erythrocyte transport of ions via the ouabain-sensitive Na-K pump, the amiloride-sensitive Na-H exchanger (NHE1), the volume and chloride-dependent K-Cl cotransporter (KCC), and the charybdotoxin-sensitive Gardos channel (KCNN4). Our data reveal substantial strain-specific and sex-specific differences in both ion content and trans-membrane ion transport in mouse erythrocytes. These differences demonstrate the feasibility of identifying specific quantitative trait loci for erythroid ion transport and content in genetically standardized inbred mouse strains.
Effect of Some Navels on Properties of Cotton/Nylon66 Blend (1 : 1) Rotor Spun Yarn and Wrapper Formation: A Comparison between Rotor and Ring Spun Yarn
Authors: Yadollah Roudbari Babak , Eskandarnejad Safdar
Published: March 2013

Use of nylon/cotton blend yarn in military uniform is common and due to advantages in its fabric in comparison to 100% cotton fabrics, capabilities of military uniforms have been improved. In this study the effects of navel type on properties of (50%-50%) nylon/cotton blend yarn and wrapper formation were investigated and compared with similar ring spun yarn. Rotor spun yarn was produced on a single head laboratory rotor spinning machine with four navels (smooth, spiral, 3 grooved, and 4 grooved) and ring yarn was produced on a zinser 319 ring spinning machine. Test result showed that navel type has a significant effect on yarn strength and strength of smooth navel yarn was maximum. Elongation of a 100% cotton rotor spun yarn is more than similar ring yarn, but it was not observed in cotton/nylon blend. Yarn irregularity and imperfections varied significantly with navel type and for rotor yarns were more than the ring yarn. Navel type had significant effect on yarn hairiness but it didnot have an effect on yarn abrasion significantly.
Some Fixed Point Theorems for Prešić-Hardy-Rogers Type Contractions in Metric Spaces
Authors: Shukla Satish , Radenović Stojan , Pantelić Slaviša
Published: March 2013

We introduce some generalizations of Prešić type contractions and establish some fixed point theorems for mappings satisfying Prešić-Hardy-Rogers type contractive conditions in metric spaces. Our results generalize and extend several known results in metric spaces. Some examples are included which illustrate the cases when new results can be applied while old ones cannot.
Gene Regulation, Modulation, and Their Applications in Gene Expression Data Analysis
Authors: Flores Mario , Hsiao Tzu-Hung , Chiu Yu-Chiao , Chuang Eric Y. , Huang Yufei , Chen Yidong
Published: March 2013

Common microarray and next-generation sequencing data analysis concentrate on tumor subtype classification, marker detection, and transcriptional regulation discovery during biological processes by exploring the correlated gene expression patterns and their shared functions. Genetic regulatory network (GRN) based approaches have been employed in many large studies in order to scrutinize for dysregulation and potential treatment controls. In addition to gene regulation and network construction, the concept of the network modulator that has significant systemic impact has been proposed, and detection algorithms have been developed in past years. Here we provide a unified mathematic description of these methods, followed with a brief survey of these modulator identification algorithms. As an early attempt to extend the concept to new RNA regulation mechanism, competitive endogenous RNA (ceRNA), into a modulator framework, we provide two applications to illustrate the network construction, modulation effect, and the preliminary finding from these networks. Those methods we surveyed and developed are used to dissect the regulated network under different modulators. Not limit to these, the concept of “modulation” can adapt to various biological mechanisms to discover the novel gene regulation mechanisms.
A Generalized If-Then-Else Operator for the Representation of Multi-Output Functions
Authors: Levin Ilya , Keren Osnat
Published: March 2013

The paper deals with fundamentals of systems of Boolean functions called multi-output functions (MOFs). A new approach to representing MOFs is introduced based on a Generalized If-Then-Else (GITE) function. It is shown that known operations on MOFs may be expressed by a GITE function. The GITE forms the algebra of MOFs. We use the properties of this algebra to solve an MOF-decomposition problem. The solution provides a compact representation of MOFs.
Protein Glycation in Diabetes as Determined by Mass Spectrometry
Authors: Lapolla Annunziata , Molin Laura , Traldi Pietro
Published: March 2013

Diabetes is a common endocrine disorder characterized by hyperglycemia leading to nonenzymatic glycation of proteins, responsible for chronic complications. The development of mass spectrometric techniques able to give highly specific and reliable results in proteome field is of wide interest for physicians, giving them new tools to monitor the disease progression and the possible complications related to diabetes, as well as the effectiveness of therapeutic treatments. This paper reports and discusses some of the data pertaining protein glycation in diabetic subjects obtained by matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS). The preliminary studies carried out byin vitroprotein glycation experiments show clear differences in molecular weight of glycated and unglycated proteins. Then, the attention was focused on plasma proteins human serum albumin (HSA) and immunoglobulin G (IgG). Enzymatic degradation products ofin vitroglycated HSA were studied in order to simulate thein vivoenzymatic digestion of glycated species by the immunological system leading to the highly reactive advanced glycation end-products (AGEs) peptides. Further studies led to the evaluation of glycated Apo A-I and glycated haemoglobin levels. A different MALDI approach was employed for the identification of markers of disease in urine samples of healthy, diabetic, nephropathic, and diabetic-nephropathic subjects.
On-Board Computing for Structural Health Monitoring with Smart Wireless Sensors by Modal Identification Using Hilbert-Huang Transform
Authors: Wu Ning , Liu Chengyin , Guo Yukun , Zhang Jianhua
Published: March 2013

Smart wireless sensors have been recognized as a promising technology to overcome many inherent difficulties and limitations associated with traditional wired structural health monitoring (SHM) systems. Despite the advances in smart sensor technologies, on-board computing capability of smart sensors has been considered as one of the most difficult challenges in the application of the smart sensors in SHM. Taking the advantage of recent developments in microprocessor which provides powerful on-board computing functionality for smart sensors, this paper presents a new decentralized data processing approach for modal identification using the Hilbert-Huang transform (HHT) algorithm, which is based on signal decomposition technique. It is shown that this method is suitable for implementation in the intrinsically distributed computing environment found in wireless smart sensor networks (WSSNs). The HHT-based decentralized data processing is, then, programmed and implemented on the Crossbow IRIS mote sensor platform. The effectiveness of the proposed techniques is demonstrated through a set of numerical studies and experimental validations on an in-house cable-stayed bridge model in terms of the accuracy of identified dynamic properties.
ALRD: AoA Localization with RSSI Differences of Directional Antennas for Wireless Sensor Networks
Authors: Jiang Jehn-Ruey , Lin Chih-Ming , Lin Feng-Yi , Huang Shing-Tsaan
Published: March 2013

In this paper, we fit RSSI values into a parabola function of the AoA between 0° and 90° by applying quadratic regression analysis. We also set up two-directional antennas with perpendicular orientations at the same position and fit the difference of the signal RSSI values of the two antennas into a linear function of the AoA between 0° and 90° by linear regression analysis. Based on the RSSI-fitting functions, we propose a novel localization scheme, called AoA Localization with RSSI Differences (ALRD), for a sensor node to quickly estimate its location with the help of two beacon nodes, each of which consists of two perpendicularly orientated directional antennas. We apply ALRD to a WSN in a [Formula: see text] m indoor area with two beacon nodes installed at two corners of the area. Our experiments demonstrate that the average localization error is 124 cm. We further propose two methods, named maximum-point minimum-diameter and maximum-point minimum-rectangle, to reduce localization errors by gathering more beacon signals within 1 s for finding the set of estimated locations of maximum density. Our results demonstrate that the two methods can reduce the average localization error by a factor of about 29%, to 89 cm.
Deoxycholic Acid as a Modifier of the Permeation of Gliclazide through the Blood Brain Barrier of a Rat
Authors: Lalić-Popović Mladena , Vasović Velibor , Milijašević Boris , Goločorbin-Kon Svetlana , Al-Salami Hani , Mikov Momir
Published: March 2013

Major problem for diabetic patients represents damage of blood vessels and the oxidative stress of the brain cells due to increased concentration of free radicals and poor nutrition of brain cells. Gliclazide has antioxidative properties and poor blood brain barrier (BBB) penetration. Bile acids are known for their hypoglycemic effect and as promoters of drug penetration across biological membranes. Accordingly, the aim of this study is to investigate whether the bile acid (deoxycholic acid) can change the permeation of gliclazide, through the blood brain barrier of a rat model type-1 diabetes. Twenty-four male Wistar rats were randomly allocated to four groups, of which, two were given alloxan intraperitoneally (100 mg/kg) to induce diabetes. One diabetic group and one healthy group were given a bolus gliclazide intra-arterially (20 mg/kg), while the other two groups apart from gliclazide got deoxycholic acid (4 mg/kg) subcutaneously. Blood samples were collected 30, 60, 150, and 240 seconds after dose, brain tissues were immediately excised and blood glucose and gliclazide concentrations were measured. Penetration of gliclazide in groups without deoxycholic acid pretreatment was increased in diabetic animals compared to healthy animals. Also in both, the healthy and diabetic animals, deoxycholic acid increased the permeation of gliclazide through that in BBB.
Triple Positive Solutions of a Nonlocal Boundary Value Problem for Singular Differential Equations withp-Laplacian
Authors: Wang Jufang , Yu Changlong , Guo Yanping
Published: March 2013

We establish the existence of triple positive solutions of anm-point boundary value problem for the nonlinear singular second-order differential equations of mixed type with ap-Laplacian operator by Leggett-William fixed point theorem. At last, we give an example to demonstrate the use of the main result of this paper. The conclusions in this paper essentially extend and improve the known results.
Spruelike Enteropathy Associated with Olmesartan: An Unusual Case of Severe Diarrhea
Authors: Dreifuss Stephanie E. , Tomizawa Yutaka , Farber Nicholas J. , Davison Jon M. , Sohnen Adam E.
Published: March 2013

A 64-year-old male with a history of hypertension presented with worsening diarrhea and 25-pound weight loss over the preceding three months. Prior screening colonoscopy was unremarkable, and the patient failed conservative management. On presentation, the patient had orthostatic hypotension associated with prerenal azotemia for which olmesartan (40 mg/day) was held. Initial workup for chronic diarrhea was essentially unremarkable. Then, EGD was performed with small bowel biopsy, which showed a moderate villous blunting and an intraepithelial lymphocyte infiltration. Celiac disease was excluded by negative conventional serology tests and the absence of clinical response to a gluten-free diet. In the interim, diarrhea became resolving without any other interventions, and clinical response was achieved even with gluten-containing diet. Two months later, he achieved a complete resolution of diarrhea and regained 20-pound weight. Spruelike enteropathy is a clinical entity manifested by chronic diarrhea and intestinal villous atrophy. Spruelike enteropathy associated with olmesartan as a cause of drug-induced diarrhea is rare, and it has been reported only in a case series to date. This case highlighted the importance for clinicians to maintain a high index of suspicion for olmesartan as a precipitant of spruelike enteropathy.
Subamolide B Isolated from Medicinal PlantCinnamomum subaveniumInduces Cytotoxicity in Human Cutaneous Squamous Cell Carcinoma Cells through Mitochondrial and CHOP-Dependent Cell Death Pathways
Authors: Yang Shu-Yi , Wang Hui-Min , Wu Tai-Wen , Chen Yi-Ju , Shieh Jeng-Jer , Lin Ju-Hwa , Ho Tsing-Fen , Luo Ren-Jie , Chen Chung-Yi , Chang Chia-Che
Published: March 2013

Subamolide B is a butanolide isolated fromCinnamomum subavenium, a medicinal plant traditionally used to treat various ailments including carcinomatous swelling. We herein reported for the first time that subamolide B potently induced cytotoxicity against diverse human skin cancer cell lines while sparing nonmalignant cells. Mechanistic studies on human cutaneous squamous cell carcinoma (SCC) cell line SCC12 highlighted the involvement of apoptosis in subamolide B-induced cytotoxicity, as evidenced by the activation of caspases-8, -9, -4, and -3, the increase in annexin V-positive population, and the partial restoration of cell viability by cotreatment with the pan-caspase inhibitor z-VAD-fmk. Additionally, subamolide B evoked cell death pathways mediated by FasL/Fas, mitochondria, and endoplasmic reticulum (ER) stress, as supported by subamolide B-induced FasL upregulation, BCL-2 suppression/cytosolic release of cytochrome c, and UPR activation/CHOP upregulation, respectively. Noteworthy, ectopic expression of c-FLIPLor dominant-negative mutant of FADD failed to impair subamolide B-induced cytotoxicity, whereas BCL-2 overexpression or CHOP depletion greatly rescued subamolide B-stimulated cells. Collectively, these results underscored the central role of mitochondrial and CHOP-mediated cell death pathways in subamolide B-induced cytotoxicity. Our findings further implicate the potential of subamolide B for cutaneous SCC therapy or as a lead compound for developing novel chemotherapeutic agents.
Split-and-Combine Singular Value Decomposition for Large-Scale Matrix
Authors: Tzeng Jengnan
Published: March 2013

The singular value decomposition (SVD) is a fundamental matrix decomposition in linear algebra. It is widely applied in many modern techniques, for example, high- dimensional data visualization, dimension reduction, data mining, latent semantic analysis, and so forth. Although the SVD plays an essential role in these fields, its apparent weakness is the order three computational cost. This order three computational cost makes many modern applications infeasible, especially when the scale of the data is huge and growing. Therefore, it is imperative to develop a fast SVD method in modern era. If the rank of matrix is much smaller than the matrix size, there are already some fast SVD approaches. In this paper, we focus on this case but with the additional condition that the data is considerably huge to be stored as a matrix form. We will demonstrate that this fast SVD result is sufficiently accurate, and most importantly it can be derived immediately. Using this fast method, many infeasible modern techniques based on the SVD will become viable.
Antidiabetic Effect and Mode of Action of Cytopiloyne
Authors: Chang Cicero Lee-Tian , Liu Hsien-Yueh , Kuo Tien-Fen , Hsu Yi-Jou , Shen Ming-Yi , Pan Chien-Yuan , Yang Wen-Chin
Published: March 2013

Cytopiloyne was identified as a novel polyacetylenic compound. However, its antidiabetic properties are poorly understood. The aim of the present study was to investigate the anti-diabetic effect and mode of action of cytopiloyne on type 2 diabetes (T2D). We first evaluated the therapeutic effect of cytopiloyne on T2D in db/db mice. We found that one dose of cytopiloyne reduced postprandial glucose levels while increasing blood insulin levels. Accordingly, long-term treatment with cytopiloyne reduced postprandial blood glucose levels, increased blood insulin, improved glucose tolerance, suppressed the level of glycosylated hemoglobinA1c(HbA1c), and protected pancreatic islets in db/db mice. Next, we studied the anti-diabetic mechanism of action of cytopiloyne. We showed that cytopiloyne failed to decrease blood glucose in streptozocin- (STZ-)treated mice whoseβcells were already destroyed. Additionally, cytopiloyne dose dependently increased insulin secretion and expression inβcells. The increase of insulin secretion/expression of cytopiloyne was regulated by protein kinase Cα(PKCα) and its activators, calcium, and diacylglycerol (DAG). Overall, our data suggest that cytopiloyne treats T2D via regulation of insulin production involving the calcium/DAG/PKCαcascade inβcells. These data thus identify the molecular mechanism of action of cytopiloyne and prove its therapeutic potential in T2D.
Stackelberg Game for Product Renewal in Supply Chain
Authors: Luo Yong , Lou Yamin , Wang Shizhao
Published: March 2013

The paper studied the process of product renewal in a supply chain, which is composed of one manufacturer and one retailer. There are original product and renewal product in the supply chain. A market share shift model for renewal product was firstly built on a increment function and a shift function. Based on the model, the decision-making plane consisting of two variables was divided into four areas. Since the process of product renewal was divided into two stages, Stackelberg-Nash game model and Stackelberg-merger game model could be built to describe this process. The optimal solutions of product pricing strategy of two games were obtained. The relationships between renewal rate, cost, pricing strategy, and profits were got by numerical simulation. Some insights were obtained from this paper. Higher renewal rate will make participants’ profits and total profit increase at the same margin cost. What is more important, the way of the optimal decision making of the SC was that RP comes onto the market with a great price differential between OP and RP.
The Missing Heritability in T1D and Potential New Targets for Prevention
Authors: Pierce Brian G. , Eberwine Ryan , Noble Janelle A. , Habib Michael , Shulha Hennady P. , Weng Zhiping , Blankenhorn Elizabeth P. , Mordes John P.
Published: March 2013

Type 1 diabetes (T1D) is a T cell-mediated disease. It is strongly associated with susceptibility haplotypes within the major histocompatibility complex, but this association accounts for an estimated 50% of susceptibility. Other studies have identified as many as 50 additional susceptibility loci, but the effect of most is very modest (odds ratio (OR) <1.5). What accounts for the “missing heritability” is unknown and is often attributed to environmental factors. Here we review new data on the cognate ligand of MHC molecules, the T cell receptor (TCR). In rats, we found that one allele of a TCR variable gene, Vβ13A, is strongly associated with T1D (OR >5) and that deletion of Vβ13+ T cells prevents diabetes. A role for the TCR is also suspected in NOD mice, but TCR regions have not been associated with human T1D. To investigate this disparity, we tested the hypothesisin silicothat previous studies of human T1D genetics were underpowered to detect MHC-contingent TCR susceptibility. We show that stratifying by MHC markedly increases statistical power to detect potential TCR susceptibility alleles. We suggest that the TCR regions are viable candidates for T1D susceptibility genes, could account for “missing heritability,” and could be targets for prevention.
Molecular Cloning, Modeling, and Characterization of Type 2 Metallothionein from Plantago ovata Forsk
Authors: Moulick Amitava , Mukhopadhyay Debashis , Talapatra Shonima , Ghoshal Nirmalya , Sen Raychaudhuri Sarmistha
Published: March 2013

Plantago ovata Forsk is a medicinally important plant. Metallothioneins are cysteine rich proteins involved in the detoxification of heavy metals. Molecular cloning and modeling of MT from P. ovata is not reported yet. The present investigation will describe the isolation, structure prediction, characterization, and expression under copper stress of type 2 metallothionein (MT2) from this species. The gene of the protein comprises three exons and two introns. The deduced protein sequence contains 81 amino acids with a calculated molecular weight of about 8.1 kDa and a theoretical pI value of 4.77. The transcript level of this protein was increased in response to copper stress. Homology modeling was used to construct a three-dimensional structure of P. ovata MT2. The 3D structure model of P. ovata MT2 will provide a significant clue for further structural and functional study of this protein.
Modified Bézier Curves with Shape-Preserving Characteristics Using Differential Evolution Optimization Algorithm
Authors: Zaman Mohammad Asif , Chowdhury Shuvro
Published: March 2013

A parametric equation for a modified Bézier curve is proposed for curve fitting applications. The proposed equation contains shaping parameters to adjust the shape of the fitted curve. This flexibility of shape control is expected to produce a curve which is capable of following any sets of discrete data points. A Differential Evolution (DE) optimization based technique is proposed to find the optimum value of these shaping parameters. The optimality of the fitted curve is defined in terms of some proposed cost parameters. These parameters are defined based on sum of squares errors. Numerical results are presented highlighting the effectiveness of the proposed curves compared with conventional Bézier curves. From the obtained
results, it is observed that the proposed method produces a curve that fits the data points more accurately.
Health-Related Quality of Life in Obese Presurgery Patients with and without Binge Eating Disorder, and Subdiagnostic Binge Eating Disorders
Authors: Sandberg Rita Marie , Dahl Jens K. , Vedul-Kjelsås Einar , Engum Bjørnar , Kulseng Bård , Mårvik Ronald , Eriksen Lasse
Published: March 2013

Objective. To study health-related quality of life (HRQoL) in obese presurgery patients with binge eating disorder (BED) and with subdiagnostic binge eating disorder (SBED) compared to patients without eating disorders or SBED.Method. Participants were patients referred to St. Olavs University Hospital, Norway, for bariatric surgery. Eating Disorders in Obesity (EDO) questionnaire was used to diagnose BED and SBED. Short-Form Health Survey (SF-12) assessed health-related quality of life. Questionnaires were returned by 160 of 209 patients. The present study sample consisted of 143 patients (103 women and 40 men) as 17 patients did not complete the SF-12.Results. Patients with BED and patients with SBED both had significantly lower mental HRQoL, but not physical HRQoL, compared to patients without eating disorders.Discussion. The findings indicate that obese presurgery patients with BED, and also SBED, may have special treatment needs in regard to their mental health.
Stability and Hopf Bifurcation Analysis of Coupled Optoelectronic Feedback Loops
Authors: Zhu Gang , Wei Junjie
Published: March 2013

The dynamics of a coupled optoelectronic feedback loops are investigated. Depending on the coupling parameters and the feedback strength, the system exhibits synchronized asymptotically stable equilibrium and Hopf bifurcation. Employing the center manifold theorem and normal form method introduced by Hassard et al. (1981), we give an algorithm for determining the Hopf bifurcation properties.
Simultaneous-Fault Diagnosis of Automotive Engine Ignition Systems Using Prior Domain Knowledge and Relevance Vector Machine
Authors: Vong Chi-Man , Wong Pak-Kin , Ip Weng-Fai , Chiu Chi-Chong
Published: March 2013

Engine ignition patterns can be analyzed to identify the engine fault according to both the specific prior domain knowledge and the shape features of the patterns. One of the challenges in ignition system diagnosis is that more than one fault may appear at a time. This kind of problem refers to simultaneous-fault diagnosis. Another challenge is the acquisition of a large amount of costly simultaneous-fault ignition patterns for constructing the diagnostic system because the number of the training patterns depends on the combination of different single faults. The above problems could be resolved by the proposed framework combining feature extraction, probabilistic classification, and decision threshold optimization. With the proposed framework, the features of the single faults in a simultaneous-fault pattern are extracted and then detected using a new probabilistic classifier, namely, pairwise coupling relevance vector machine, which is trained with single-fault patterns only. Therefore, the training dataset of simultaneous-fault patterns is not necessary. Experimental results show that the proposed framework performs well for both single-fault and simultaneous-fault diagnoses and is superior to the existing approach.
Abstract A15: Snail upregulation of SPARC leads to increased invasion in models of both early-and late-stage NSCLC
Authors: Grant Jeanette L. , Walser Tonya C. , Minna John D. , Shay Jerry W. , Dubinett Steven M.
Published: March 2013

The zinc-finger protein Snail is upregulated in human non-small cell lung cancer (NSCLC) tissues and is associated with poor patient prognosis. We have shown that Snail upregulation in lung cancer cell lines leads to morphologic changes indicative of epithelial-mesenchymal transition (EMT) in vitro and tumor progression in vivo. Snail overexpression is associated with differential gene expression related to diverse aspects of lung cancer progression. One gene upregulated by Snail in both NSCLC and histologically normal human bronchial epithelial cell lines is secreted protein, acidic and rich in cysteine (SPARC). Expression of SPARC modulates reversible interactions between cells and their extracellular matrix and is known to play an important role in the wound healing process of normal epithelial cells. Its upregulation in several cancer types is associated with increased migration, invasion, and poor patient prognosis. Here, we show that overexpression of Snail leads to increased invasion in a modified Boyden chamber assay in models of both early and late stage NSCLC. Knockdown of SPARC by shRNA leads to a reversal of invasion, indicating that SPARC is at least partially responsible for the effect of Snail on invasion. Computational analysis indicates that Snail does not directly enhance the transcription of SPARC; therefore, we have investigated the indirect mechanism for this relationship. Both ERK1/2 and the chaperone protein HSP27 have emerged as potential components of the pathway. Snail overexpression also leads to activation of ERK1/2, and chemical inhibition of this pathway leads to a decrease in SPARC protein. Snail-overexpressing cell lines show increased transcription of HSP27 mRNA. As Snail has been shown to be overexpressed in the inflammatory microenvironment of premalignancy and in established tumors, SPARC-driven invasion supports both the parallel and linear models of tumor progression. Delineating pathways involved in Snail-dependent and SPARC-mediated tumor progression may yield new targets for lung cancer prevention and treatment.
Abstract A19: Lung cancers taken from autopsies express more tumor antigens than those taken from surgical resections: Implications for immunoprevention
Authors: Ge Lisheng , Lambrecht Nils , Ouyang Yi , Hoa Neil , Chen Longchaun , Deng Xiamong , Dacosta-Iyer Maria , Szabo Sandor , Jadus Martin Robert
Published: March 2013

Lung cancers are fatal cancers, even if caught early, since some cancer cells have spread by the time they are detected. Some lung cancer patients treated by immunotherapy have improved their survival. A limited number of tumor-associated antigens (TA), both humoral-based and cell-mediated antigens, are known for lung cancers. The identification of more relevant antigens will allow better immunotherapies to be developed. We analyzed adenocarcinomas and small cell lung cancers taken from surgical resections or from autopsies to determine the prevalent antigens by using quantitative real time PCR techniques for 43 putative tumor antigens. We compared these tumor-derived profiles with normal lung cell expression. The data fell into 3 profiles where: 1) both tumors were positive for 18 TA; 2) only SCLC expressed 14 TA; 3) a group of 10 TA that had no differences. The lung tumor samples taken at autopsy, expressed a higher number of TA and expressed more mRNA TA than those taken from surgical resections. Blotting techniques using the specimens from the lung cancer autopsies confirmed increased expression of the tumor antigen precursor proteins when compared to the normal lung. Use of two lung cancer cell lines also independently confirmed the presence of these proteins. These data suggest that human lung cancers at the time of death are more antigenic than those taken at an earlier stage. This type of tumor taken at autopsy might provide more targets for vaccine generation and allow the concept of immunoprevention to be used in lung cancer patients.
Abstract A21: Tissue platinum concentration and tumor response in non-small cell lung cancer
Authors: Kim Eric S. , He Guangan , Chow Chi-Wan , Fujimoto Junya , Lee Jack J. , Kalhor Neda , Swisher Stephen G. , Wistuba Ignacio I. , Stewart David J. , Siddik Zahid H.
Published: March 2013

Background: Platinum resistance is a major limitation in the treatment of advanced non-small cell lung cancer (NSCLC). Reduced intracellular drug accumulation is one of the most consistently identified features of platinum-resistant cell lines, but clinical data are limited. We assessed effects of tissue platinum concentrations on tumor response, time to recurrence, progression-free survival (PFS) and overall survival (OS) in patients with early-stage NSCLC following neoadjuvant platinum-based chemotherapy.

Methods: We measured total platinum concentrations in 44 archived fresh frozen NSCLC specimens from patients who received neoadjuvant platinum-based chemotherapy. Samples were analyzed by flameless atomic absorption spectrophotometry to assess absorbance reading associated with platinum concentration. Four specimens from patients who underwent surgery only and one patient who received pemetrexed as neoadjuvant chemotherapy were analyzed as negative controls. Absorbance value per mg of tissue was correlated with percent reduction in tumor size on post- vs prechemotherapy CT scans. Kaplan-Meier curves and log-rank tests were used to evaluate differences in time to recurrence and survival between two groups divided by median platinum concentration.

Results: Platinum absorbance values in 44 neoadjuvant specimens were easily detectable and ranged from 0.6 to 6.7 × 10−3 per mg of tissue while five negative controls demonstrated absorbance readings similar to 0.1N HCL. Platinum absorbance correlated significantly with percent reduction in tumor size (R2 = 0.48, P<0.00001). The same correlations were seen in cisplatin (P = 0.0003), carboplatin (P = 0.0013), adenocarcinoma (P = 0.0003) and squamous cell carcinoma (P = 0.019) groups. Furthermore, there was no significant impact of potential variables such as the type of platinum compound, number of cycles, pre-treatment tumor diameter and time lapse from last chemotherapy on platinum concentration. Patients with lower Pt concentration also had shorter time to recurrence (P = 0.0494), progression free survival (P = 0.0366) and overall survival (P = 0.035).

Conclusion: This is the first tissue-based study to establish a relationship between tissue platinum concentrations and response in NSCLC. Reduced intratumoral platinum accumulation may constitute a significant mechanism of platinum resistance even in clinical specimens. Further studies investigating factors that modulate intracellular platinum concentration are warranted (Supported by National Foundation of Cancer Research 90088436, Department of Defense W81XWH-07-1-0306, National Institute of Health CA127263, CA160687 and CA16672).
Abstract A22: Clinical implication of microscopic anthracotic pigment in mediastinal staging of non-small cell lung cancer by endobronchial ultrasound-guided transbronchial needle aspiration
Authors: Kim Young Whan , Park Young Sik , Lee Jinwoo , Lee Sang-Min , Yim Jae-Joon , Han Sung Koo , Pang Jin Chul , Chung Doo Hyun , Yang Seok-Chul
Published: March 2013

Introduction: Microscopic anthracotic pigment is frequently observed in EBUS-TBNA specimen from non-small cell lung cancer, but the clinical interpretation is not known. The aim of this study was to evaluate the clinical implication of microscopic anthracotic pigment in mediastinal staging of non-small cell lung cancer by EBUS-TBNA.

Methods: From May 2010 to July 2011, Consecutive potentially operable non-small cell lung cancer patients who underwent EBUS-TBNA for mediastinal staging were recruited in this retrospective study.

Results: Total 133 patients, male were 102 (76.7%). Median age was 68. Squamous cell carcinoma (59, 44.4%) and adenocarcinoma (57, 42.9%) were most common. Clinical stages after EBUS-TBNA were as followed; IA (18.8%), IB (17.3%), IIA (6.0%), IIB (9.0%), IIIA (36.8%), IIIB (12.0%). Total 281 mediastinal lymph nodes were sampled by EBUS-TBNA, station 4R (100, 35.8%) and station 7 (86, 30.8%) were most common sites. Malignant lymph nodes were 100 (35.8%). Microscopic anthracotic pigment was observed in 61 (21.7%) lymph nodes, and among them malignant lymph nodes were only 3 (P<0.001). The lymph nodes with microscopic anthracotic pigment were small (9.0 vs. 10.8 mm, P = 0.001) and low SUV on FDG-PET (5.4 vs. 6.7, P = 0.120). In multivariable analysis, microscopic anthracotic pigment was statistically significant associated with benign lymph node (adjusted OR, 14.3; 95% CI, 3.6–57.3; P<0.001).

Conclusion: In potentially operable non-small cell lung cancer patients, microscopic anthracotic pigment in endobronchial ultrasound-guided transbronchial needle aspiration specimen was strongly associated with benign mediastinal lymph node.
Abstract A24: CpG island methylation of microRNAs is associated with tumor size and the recurrence of non-small cell lung cancer
Authors: Kitano Kentaro , Goto Akiteru , Fukayama Masashi , Yatomi Yutaka , Ohishi Nobuya , Takai Daiya , Watanabe Kousuke , Emoto Noriko , Kage Hidenori , Hamano Emi , Nagase Takahide , Sano Atsushi , Murakawa Tomohiro , Nakajima Jun
Published: March 2013

We investigated whether the CpG island methylation of certain microRNAs was associated with the clinicopathological features and the prognosis of non-small cell lung cancer. The methylation of mir-152, -9-3, -124-1, -124-2, and -124-3 was analyzed in 96 non-small cell lung cancer specimens using a combined bisulfite restriction analysis. The median observation period was 49.5 months. The methylation of mir-9-3, -124-2, and -124-3 was individually associated with an advanced T factor independently of age, sex, and smoking habit. Moreover, the methylation of multiple microRNA loci was associated with a poorer progression-free survival in a univariate analysis. Our result enlightens the accumulation of aberrant DNA methylation which occurs in concordance with the tumor progression.
Abstract A27: Expression of the EGFR suppressor LRIG1 is associated with survival in non-small cell lung cancer
Authors: Kvarnbrink Samue , Edlund Karolina , Botling Johan , Micke Patrick , Hedman Håkan , Johansson Mikael
Published: March 2013

Leucine-rich repeats and immunoglobulin-like domains (LRIG)-1 is an endogenous inhibitor of epidermal growth factor receptor (EGFR) signaling, that is down-regulated in several forms of human cancer. The LRIG gene family also includes LRIG2 and LRIG3. EGFR signaling is important in non-small cell lung cancer (NSCLC) and EGFR tyrosine kinase inhibitors are today considered standard treatment in patients with advanced disease. Since smoking recently was reported to down-regulate LRIG1 gene expression in lung epithelium and squamous cell carcinoma of the lung, the LRIG family of proteins may be important in lung cancer.

The objective of the present study was to investigate if LRIG is a prognostic factor in non-small cell lung cancer (NSCLC). Therefore, we conducted an in silico analysis of LRIG gene expression in publicly available datasets using the Oncomine database and compared LRIG expression to clinical outcome. We also analyzed the LRIG protein expression by immunohistochemistry in a tissue micro array (TMA, n = 190) representing long- and short time survivors from a surgically treated NSCLC patient cohort.

In seven out of 11 datasets in the in silico analysis, high expression of LRIG1 correlated (p<0.05) with long patient survival. This correlation was apparent both in adenocarcinoma and squamous cell carcinoma. In the TMA, LRIG1 expression was significantly higher in the long-term survival group than in the short time survival group (p<0.05). LRIG2 and LRIG3 protein expression did not significantly correlate to survival.

In conclusion, both our in silico analysis of gene expression and TMA study of protein expression showed associations between high LRIG1 expression and longer NSCLC patient survival. This implicates that LRIG1 may be a prognostic factor in NSCLC and further studies on larger datasets have been initiated.
Abstract A29: Genomic alteration of E2F/Rb activates EZH2 in small cell lung cancer
Authors: Thu Kelsie L. , Hubaux Roland , Aviel-Ronen Sarit , MacAulay Calum E. , Tsao Ming-Sound , Lam Stephen , Lam Wan L. , Coe Bradley P.
Published: March 2013

Introduction: Small Cell Lung Cancer (SCLC) is a highly aggressive lung tumor with a 5 year survival rate of only 5% for extensive stage disease, which has only modestly improved over the last few decades. Identification of new molecular diagnostic and therapeutic targets is thus imperative. The purpose of this study was to employ genomic profiling of SCLC tumors to identify novel genomic alterations responsible for driving the aggressive biology of SCLC.

Methods: DNA was extracted from a panel of 14 formalin fixed paraffin embedded SCLC tumors and used to perform high resolution array comparative genomic hybridization (aCGH) on a tiling array to identify recurrent copy number alterations. RNA was harvested from a series of SCLC cell lines and used to perform qRT-PCR on candidate genes. Publicly available SCLC tumor microarray data was also analyzed to interrogate the expression of candidate genes.

Results: Through application of integrated genome and transcriptome microarray profiling, and comparison of SCLC to less aggressive non-small cell lung cancers (NSCLC) we have identified novel patterns of genomic alteration mediated pathway disruption specific to SCLC. This includes activation of the cell cycle through deregulation of downstream pathway components, as opposed to upstream deregulation of receptors such as EGFR which is characteristic of NSCLC.

Strikingly we observed direct genomic activation of E2F transcription factors, in addition to the classically described loss of the Rb tumor suppressor. Analysis of targets of the E2F/Rb pathway identified EZH2, a polycomb repressive complex 2 (PRC2) member involved in epigenetic silencing of genes involved in differentiation. EZH2 has been characterized as a target of genomic amplification in prostate and breast cancers, however, no genomic amplifications were detected in our SCLC samples, thus any overexpression is likely regulated by upstream elements of the E2F/Rb pathway. qRT-PCR confirmed EZH2 as being specifically hyper-activated with a mean 42 fold over-expression (range 10 to 74 fold) in SCLC compared to 13 fold in NSCLC. This pattern was verified from an analysis of an independent array study of SCLC. Moreover, shRNA-mediated knockdown demonstrated a significant reduction in cell viability in SCLC cell lines.

Conclusion: We conclude that EZH2 activation through genomic alteration of E2F/Rb contributes to the malignant phenotype of SCLC and may represent a potential therapeutic target.
Abstract A33: Markov model of VeriStrat© selection criteria for EGFR Inhibitor therapy predicts improved survival for previously treated patients with NSCLC
Authors: Nelson Richard E. , Akerley Wallace
Published: March 2013

Background: EGFR inhibitor therapy extends survival as 2nd or 3rd line treatment of patients with previously treated metastatic non-small cell lung cancer (NSCLC), but a randomized trial of 2nd line therapy in unselected patients showed similar benefit versus conventional chemotherapy. EGFR mutation analysis is useful in choosing therapy, but it is applicable to only a minority of patients due to low frequency of mutation in a North American population and ignores the majority of patients who may benefit with wild type status. Alternative selection strategies are needed. VeriStrat is a protein expression profile utilizing matrix-assisted laser desorption/ionization time of flight mass spectrometry to analyze serum. The test classifies patients as VeriStrat Good or VeriStrat Poor based on likelihood of benefitting from EGFR inhibitor therapy. The objective of this research was to model the anticipated survival of patients using a Veristrat Selection Strategy for 2nd line therapy of NSCLC to the alternative treatment strategies of chemotherapy for all (Chemoall), EGFR inhibitor to all (EGFR-all) l, or to performance status selection (PS selection), a common practice, where those with good performance status receive chemotherapy and those with poor PS receive EGFR inhibitor therapy.

Methods: We developed a Markov model to estimate the expected survival of patients for 4 different treatment strategies for NSCLC: (1) VeriStrat Selection, (2) chemotherapy to all patients, (3) EGFR inhibitor to all patients, and (4) PS selection. Model inputs, taken from published literature, for the base-case analysis (and ranges) were the following: PS 0-1, 50% (35%–65%); VeriStrat Good, 60% (45%–75%); VeriStrat Good if PS >1, 40% (25%–55%); 1-year survival on EGFR inhibitor if VeriStrat Good, 47% (43–50); 1-year survival on EGFR inhibitor if VeriStrat Poor, 12% (8–16) and 1-year survival on chemotherapy, 29% (23–35). Probabilistic sensitivity analysis was performed using 10,000 2nd order Monte Carlo simulations.

Results: In the base case analysis, the median survival for EGFR-all was 8.97 months, Chemo-all was 8.48 months, PS selection was 8.20 months and VeriStrat selection was 10.03 months. In the sensitivity analysis, the VeriStrat selection strategy yielded the longest survival in each of the 10,000 simulations.

Conclusion: These results suggest that VeriStrat-based selection of “standard of care” 2nd line therapy of NSCLC improves survival to a clinically significant degree. This strategy, in the absence of new therapies, appears to improve median survivals by 4–6 weeks.
Abstract A38: Whole methylome explorations of paired lung tumor and non-tumor clinical samples
Authors: Mullapudi Nandita , Suzuki Masako , Fazzari Melissa , Han Weiguo , Spivack Simon D.
Published: March 2013

Background: Hypermethylation in specific candidate gene promoters has been found during progressive lung carcinogenesis. To explore common methylation events on a genome-wide scale in lung cancer, we analyzed the methylation profiles of paired NSCLC tumor and far adjacent non-tumor samples using the HELP-microarray assay, which yields information on 1.2 million fragments throughout the genome.

Methods: The HELP (HpaII tiny fragment Enriched by Ligation mediated PCR) assay is based on the generation of restriction enzyme libraries generated by methylation sensitive (HpaII) and methylation insensitive (MspI) isoschizomers, in its second generation as a microarray platform. The assay lends itself to low starting amounts of DNA (3 ug) and robust assessment of methylation status by comparing ratios of HpaII- generated-fragments to MspI- generated fragments co-hybridized to a Nimbelgen custom high-density microarrays. The CCGG sites were weighted if neighboring CCGGs were methylated in same direction. Here, 24 pairs of tumor and adjacent non-tumor samples were analyzed using the HELP assay.

Summary of results: At p = 5E-6, we identified 26,138 differentially methylated fragments (corresponding to 2 CpG sites each) in tumor versus non-tumor. The overall trend was consistent with genome-wide hypomethylation and locus specific hypermethylation (localized to CG-island containing promoters). We could identify both known and novel regions of the genome as well as specific gene-promoters that are hypermethylated in tumor versus non-tumor.

Region # loci # signif loci T Hypomethyl T Hypermethyl

Promoter 151,568 576 69% 31%

Gene Body 551,628 9,817 62% 38%

Intergenic 540,473 15,745 97% 3%

Conclusion: An interrogation of methylation status of 1.2 million loci throughout the genome in paired lung tumor/non-tumor specimens reveals many more differential methylation events in gene bodies and intergenic regions than in promoters. That said, many previously unreported differentially-methylated gene promoters were identified. We are able to discover individual methylation events common across different clinical specimens. Based on a set of priors, we have narrowed down promoter-specific hypermethylation events for further validation using tagged Bisulfite Genomic Sequencing. We are also working on the integration of methylome date with other genome-wide epigenetic and expression data from the same clinical samples.

[Funding source: NCI 1RC1 CA145422-01; 1K24-CA139054-01]