bluuebunny/crossref_metadata_2025 huggingface dataset:
Racial disparities and survival for nonsmall‐cell lung cancer in a large cohort of black and white elderly patients
Authors: Hardy Dale , Xia Rui , Liu Chih‐Chin , Cormier Janice N. , Nurgalieva Zhannat , Du Xianglin L.
Published: July 2009

BACKGROUND:

This study aimed to examine disparities in survival and associated factors for patients with nonsmall‐cell lung cancer (NSCLC) and to determine whether racial disparities varied over time (1991‐1995, 1996‐1999, and 2000‐2002).

METHODS:

The authors studied 70,901 patients aged ≥65 years with stage I‐IV NSCLC identified from Surveillance, Epidemiology, and End Results/Medicare data. Multivariate time‐to‐event survival analyses were completed using Cox proportional regression modeling.

RESULTS:

The 5‐year observed lung cancer‐specific survival rates were 52.7% for whites and 47.5% for blacks with stage I‐II disease, and 17.7% and 19.6% for whites and blacks, respectively at stages III‐IV. After controlling for standard treatment, socioeconomic status (SES), and other factors, there were no significant differences in all‐cause mortality, or lung cancer‐specific mortality between black and white patients with stage I‐II or III‐IV lung cancer. However, blacks had an increased risk for overall all‐cause mortality at stage I‐IV (hazard ratio [HR], 1.24; 95% confidence interval, 1.13‐1.35), and during 2000‐2002 at stage III‐IV for all‐cause mortality (HR, 1.22; 95% CI, 1.02‐1.47) and lung cancer‐specific mortality (HR, 1.24; 95% CI,1.01‐1.53). Standard treatment was significantly associated with increased survival, whereas poor SES was associated with increased mortality.

CONCLUSIONS:

There were no significant differences in survival between blacks and whites with NSCLC within stage stratifications after adjusting for covariates, except for black patients at overall stage for all‐cause mortality and at stage III‐IV diagnosed in 2000‐2002. Receiving stage‐specific evidence‐based standard therapy was associated with significantly increased survival. Cancer 2009. © 2009 American Cancer Society.
Parallelizing dense and banded linear algebra libraries using SMPSs
Authors: Badia Rosa M. , Herrero José R. , Labarta Jesús , Pérez Josep M. , Quintana‐Ortí Enrique S. , Quintana‐Ortí Gregorio
Published: July 2009

The promise of future many‐core processors, with hundreds of threads running concurrently, has led the developers of linear algebra libraries to rethink their design in order to extract more parallelism, further exploit data locality, attain better load balance, and pay careful attention to the critical path of computation. In this paper we describe how existing serial libraries such as (C)LAPACK and FLAME can be easily parallelized using the SMPSs tools, consisting of a few OpenMP‐like pragmas and a run‐time system. In the LAPACK case, this usually requires the development of blocked algorithms for simple BLAS‐level operations, which expose concurrency at a finer grain. For better performance, our experimental results indicate that column‐major order, as employed by this library, needs to be abandoned in benefit of a block data layout. This will require a deeper rewrite of LAPACK or, alternatively, a dynamic conversion of the storage pattern at run‐time. The parallelization of FLAME routines using SMPSs is simpler as this library includes blocked algorithms (or algorithms‐by‐blocks in the FLAME argot) for most operations and storage‐by‐blocks (or block data layout) is already in place. Copyright © 2009 John Wiley & Sons, Ltd.
The role of cervical smear in the diagnosis and management of extrauterine malignancies metastatic to the cervix: Three case reports
Authors: Giordano Giovanna , Gnetti Letizia , Pilato Francesco Paolo , Viviano Luca , Silini Enrico Maria
Published: July 2009

This report analyses the clinicopathologic features of three metastatic extragenital neoplasms to the cervix. These examples of metastatic extragenital malignancies to the cervix are cutaneous malignant melanoma, colorectal carcinoma, and breast carcinoma. The diagnosis of these metastatic malignancies was suspected on a historical basis. The value of Pap smears is limited on the other hand, since for a correct diagnosis in all our cases, this examination required corroboration by histological and immunohistochemical analysis. In all these examples of extragenital metastatic neoplasms, as in other cases reported in literature, the patients had undergone cervical smear because of vaginal bleeding, even if Pap smear is considered a questionable maneuver in vaginal bleeding. Accordingly, our article reveals that a great deal of rigorous screening, precise morphological analysis, and plentiful clinical data are mandatory in approaching diagnoses of extracervical malignancies. However, this study did demonstrate that cytology does not alter patient management or outcome. In effect, in accordance with other studies, our cases of metastatic extragenital neoplasms to the uterus have poor prognosis Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.
Ricin poisoning and forensic toxicology
Authors: Musshoff Frank , Madea Burkhard
Published: July 2009

Ricin is one of the most fascinating poisons due to its high toxicity: as little as 500 µg can kill an adult. It gained fame by its use in the so‐called ‘umbrella murder’ to kill the Bulgarian dissident Georgi Markov in 1978. Ricin also became known as a potential bio‐terror agent to which people could be exposed through the air, food, or water. The origin, biochemistry, toxicity, and analytical procedures for the determination of ricin are summarized. The homicide of Markov is described as well as recent cases of criminal ricin use. Copyright © 2009 John Wiley & Sons, Ltd.
Murder by insulin: suspected, purported and proven—a review
Authors: Marks Vincent
Published: July 2009

Murder by insulin—whether attempted, suspected or proven—is rare. Only 66 cases worldwide could be found for this review. A conviction was secured in 31 cases and additional weapon was employed in 11. Differentiation of attempted homicide from Munchausen syndrome by proxy in the young and from ‘mercy killing’ in the elderly was not attempted. Most perpetrators were close relatives and most victims were alive when discovered and responded to treatment. Hypoglycaemia is the first clue to homicidal insulin use in living subjects and requires the demonstration of a plasma insulin concentration of generally more than 1000 pmol/L and undetectable plasma C‐peptide concentration to establish the diagnosis. Serum glucose measurements are valueless in victims found dead. The presence near the body of insulin vials, syringes or needles, loose talk by the suspected perpetrator or their ready access to insulin may be the only clue. The demonstration of insulin in tissue around an injection site by immunohistopathology or by measuring it in an extract clinches the diagnosis. Immunoassays suitable for clinical use to detect and measure insulin and C‐peptide are subject to random errors and cannot be relied upon unless special precautions including separation by gel filtration or HPLC are undertaken prior to analysis. They do not detect or measure accurately a new generation of synthetic insulin analogues. Mass spectrometry will be required to do this and to validate clinical immunoassays, upon which convictions have always had to rely in the past. Copyright © 2009 John Wiley & Sons, Ltd.
Multi target analysis of putrefactive specimens by liquid chromatography—tandem mass spectrometry to prove multiple poisonings by hypnotics and muscle relaxants
Authors: Thieme Detlef , Peschel Oliver , Fischer Florian , Graw Matthias
Published: July 2009

The muscle relaxant rocuronium and the hypnotics etomidate, diazepam and midazolam were candidate poisons in a case of a suspected series of fatal intoxications in a hospital. A robust, specific and sensitive multi‐target screening procedure was established to identify unequivocally the parent compounds and diagnostic metabolites and artefacts in putrefied specimens, obtained from exhumed bodies. The analytical findings of relevant compounds could be traced partially back to authorised therapeutic measures, whereas the identification of rocuronium proved potentially lethal intoxications in 13 (of a total number of 42) cases.

Moreover, the detection of certain hypnotics revealed an improper administration of these compounds in another nine cases, which suggested manipulation but was not indicative of fatal intoxications.

Quantitative estimations of substance concentrations were highly correlated with the post‐mortem time intervals and did not reveal any information on doses, initial serum concentrations or toxicological effects. Copyright © 2009 John Wiley & Sons, Ltd.
Murder by poisoning: successful analytical investigations of spectacular cases in Austria
Authors: Vycudilik Walter , Gmeiner Günter
Published: July 2009

To prove murder by poisoning requires the application of analytical toxicology to detect the fatal substance and clear up the cause of death. Improvements in the development of mass spectrometry in combination with high‐resolution chromatographic methods are steadily enhancing detection and identification power but making use of these advances relies on proper sample preparation as well as on knowledge about the chemical nature of the substances and their bio‐transformation products. This review gives examples of case reports with successful analytical investigations of murder by poisoning in spectacular Austrian cases involving low molecular weight. Copyright © 2009 John Wiley & Sons, Ltd.
Synthesis, Crystal Structures and NMR Spectroscopic Studies on Ruthenium Complexes with Phosphanylacetal and Phosphanylthioacetal Ligands
Authors: Bayly Simon R. , Cowley Andrew R. , Dilworth Jonathan R. , Ward Caroline V.
Published: July 2009

The chemistry of the mixed P,S‐donor ligands [2‐(1,3‐dioxolan‐2‐yl)phenyl]diphenylphosphane (PhPOO, 1), and [2‐(1,3‐dithiolan‐2‐yl)phenyl]diphenylphosphane (PhPSS, 2) with the RuII precursors [RuCl2(PPh3)3] and [RuCl2(4‐cymene)]2 has been investigated. The structures of the resulting complexes were analysed by X‐ray crystallography and 1H, 13C and 31P NMR spectroscopy, and their activity as catalysts for hydrosilylation was examined. Reaction of 1 with [RuCl2(PPh3)3] in methanol solution produced [RuCl2(PhP(OMe)2)2] (3) {with [RuCl2(MeOH){PhP(OMe)2}(PPh3)] (3a) as a side product}, whereas the same reaction with [RuCl2(4‐cymene)]2 produced [RuCl2(PhPOO)2] (4). The dioxolane complex 4 showed fluxional behaviour by NMR spectroscopy, whereas 3 did not. Reaction of PhPSS with[RuCl2(PPh3)3] in methanol solution produced [RuCl2(PhPSS)2] (5), and reaction with [RuCl2(4‐cymene)]2 produced the cationic complex [RuCl(4‐cymene)(PhPSS)]+ (6) by precipitation with NaBPh4. In solution both 5 and 6 exist in two isomeric forms, and neither shows evidence of fluxionality at room temperature. Of the complexes tested only the acetal species 3 showed any significant hydrosilylation activity. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
A Tetramethylplatinum(IV) Complex with 1,1′‐Bis(diphenylphosphanyl)ferrocene Ligands: Reaction with Trifluoroacetic Acid
Authors: Shahsavari Hamid R. , Rashidi Mehdi , Nabavizadeh S. Masoud , Habibzadeh Sepideh , Heinemann Frank W.
Published: July 2009

A new tetramethylplatinum(IV) complex [PtMe4(dppf)] [2; dppf = 1,1′‐bis(diphenylphosphanyl)ferrocene], as the first platinum(IV) complex to contain a chelating dppf ligand, was prepared by the reaction of the known dimeric tetramethylplatinum(IV) complex cis,cis‐[Me4Pt(μ‐SMe2)2PtMe4] (1) with the biphosphane ligand dppf (2 equiv.) at room temperature by replacement of the SMe2 ligands with the P ligating atoms of dppf. The single‐crystal X‐ray structure of complex 2 revealed that the dppf chelating ligand is arranged close to the “synperiplanar–eclipsed” conformation, with a Cp(centroid)···Fe···Cp(centroid) twist angle of 17.6° and a dppf bite angle, P1–Pt1–P2, of 95.77(3)°. This is in contrast to the usually preferred “synclinal–staggered” conformation, in which the Cp(centroid)···Fe···Cp(centroid) twist angle is close to 36° as found in PtII complexes with chelating dppf ligands, like for example [PtMe2(dppf)] exhibiting a dppf bite angle close to 100°. When complex 2 was treated with the strong acid CF3COOH (1 equiv.) first evolution of methane was observed, followed by a C–C coupling reaction to give ethane and the methylplatinum(II) complex [PtMe(OCOCF3)(dppf)] (3). The reaction of complex 2 with the nucleophile HgCl2 (1 equiv.) similarly gave MeHgCl, ethane, and the methylplatinum(II) complex [PtMeCl(dppf)] (11). The structure of complex 3 was also determined by X‐ray crystallography. The geometry around the platinum center is best described as distorted square‐planar and the dppf is arranged in the usually preferred “synclinal–staggered” conformation.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Synthesis and Redox Behavior of 1,3‐Bismethylthio‐) and 1,3‐Bis(phenylthio)azulenes Bearing 2‐ and 3‐Thienyl Substituents by Palladium‐Catalyzed Cross‐Coupling Reaction of 2‐ and 6‐Haloazulenes with Thienylmagnesium Ate Complexes
Authors: Shoji Taku , Ito Shunji , Toyota Kozo , Iwamoto Takeaki , Yasunami Masafumi , Morita Noboru
Published: July 2009

Preparation of thienylazulenes 3–6 was established by the palladium‐catalyzed cross‐coupling reaction of the corresponding haloazulenes with thienylmagnesium ate complexes, which were readily prepared from the corresponding bromothiophenes. The reaction of 3–6 with several sulfoxides in the presence of Tf2O, followed by treatment with triethylamine afforded the corresponding 1,3‐bis(methylthio)‐ and 1,3‐bis(phenylthio)‐2‐ and 6‐thienylazulenes 7–12 in good yields. Redox behavior of the novel azulene derivatives with 2‐ and 3‐thienyl‐substituted 3–12 was examined by cyclic voltammetry and differential pulse voltammetry, which revealed amphoteric redox behavior. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Reactions between 1‐Ethynylazulenes and 7,7,8,8‐Tetracyanoquinodimethane (TCNQ): Preparation, Properties, and Redox Behavior of Novel Azulene‐Substituted Redox‐Active Chromophores
Authors: Shoji Taku , Ito Shunji , Toyota Kozo , Iwamoto Takeaki , Yasunami Masafumi , Morita Noboru
Published: July 2009

[2+2] Cycloaddition/cycloreversion reactions between TCNQ and mono‐ or bis[2‐(azulen‐1‐yl)ethynyl]benzene or ‐thiophene derivatives were examined: the corresponding TCNQ adducts, novel azulene‐substituted redox‐active chromophores, were produced in excellent yields. TCNE/TCNQ double adducts were also prepared both by stepwise and by one‐pot cascade reactions. The redox behavior of these novel azulene‐substituted chromophores was examined by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Significant color changes in these azulene‐substituted chromophores under electrochemical reduction conditions were observed by visible spectroscopy. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
An evaluation of a 3D free‐energy‐based lattice Boltzmann model for multiphase flows with large density ratio
Authors: Huang Haibo , Zheng Hongwei , Lu Xi‐yun , Shu Chang
Published: July 2009

In this paper, the 3D Navier–Stokes (N–S) equation and Cahn–Hilliard (C–H) equations were solved using a free‐energy‐based lattice Boltzmann (LB) model. In this model, a LB equation with a D3Q19 velocity model is used to recover continuity and N–S equations while another LB equation with D3Q7 velocity model for solving C–H equation (Int. J. Numer. Meth. Fluids, 2008; 56:1653–1671) is applied to solve the 3D C–H equation.

To avoid the excessive use of computational resources, a moving reference frame is adopted to allow long‐time simulation of a bubble rising. How to handle the inlet/outlet and moving‐wall boundary conditions are suggested. These boundary conditions are simple and easy for implementation.

This model's performance on two‐phase flows was investigated and the mass conservation of this model was evaluated. The model is validated by its application to simulate the 3D air bubble rising in viscous liquid (density ratio is 1000). Good agreement was obtained between the present numerical results and experimental results when Re is small. However, for high‐Re cases, the mass conservation seems not so good as the low‐Re case. Copyright © 2009 John Wiley & Sons, Ltd.
Detectable BRAF mutation in serum DNA samples from patients with papillary thyroid carcinomas
Authors: Chuang Tony C. Y. , Chuang Alice Y. C. , Poeta Luana , Koch Wayne M. , Califano Joseph A. , Tufano Ralph P.
Published: July 2009

Background.

An activating point mutation of the BRAF oncogene results in a V600E amino acid missense mutation found in a majority of papillary thyroid carcinomas (PTC).

Methods.

In this study, 28 matched tumor and serum samples obtained from patients with both benign and malignant thyroid disorders were analyzed for BRAF mutation using a gap‐ligase chain reaction technique.

Results.

The BRAF mutation was absent in tumor DNA samples obtained from patients with benign adenomas, follicular neoplasms or carcinoma, and thyroid lymphoma. In contrast, 5 of 14 PTC tumors were positive for the BRAF mutation. Moreover, 3 of 14 patients with PTC were positive for BRAF mutation in serum and tumor. Of these 3 patients, 2 had lymph node metastasis and 2 had PTC in background of the Hashimoto's thyroiditis.

Conclusions.

The detection of free circulating mutant BRAF in patients with PTC is possible and future studies are warranted to determine its clinical significance. © 2009 Wiley Periodicals, Inc. Head Neck, 2010
Multidimensional analysis of body image concerns among newly diagnosed patients with oral cavity cancer
Authors: Fingeret Michelle Cororve , Vidrine Damon J. , Reece Gregory P. , Gillenwater Ann M. , Gritz Ellen R.
Published: July 2009

Background.

Body image is a critical psychosocial issue for patients facing treatment for oral cancer, yet there is limited research conducted in this area. This study uses a multidimensional approach to body image assessment and evaluates relationships between body image, demographic, health, and psychosocial variables.

Methods.

Newly diagnosed patients with oral cancer completed self‐report questionnaires and a structured clinical interview.

Results.

Most participants identified current and/or future body image concerns primarily related to impending surgery. Adequate psychometric properties were demonstrated on a range of body image measures. Depression was the strongest and most consistent predictor of body image outcomes.

Conclusions.

Preliminary evidence supports the importance of evaluating body image concerns in oral cancer patients prior to surgical intervention. Our findings have implications for developing validated body image tools and can be used to guide psychosocial interventions targeting body image disturbance. © 2009 Wiley Periodicals, Inc. Head Neck, 2010
Anatomical landmarks for endosonography of the larynx
Authors: Kraft Marcel , Mende Susanne , Arnoux André , Arens Christoph
Published: July 2009

Background

A precise knowledge of anatomy is necessary to allow a correct interpretation of sonographic images when investigating a particular region of the body. The objective of the present study was to establish anatomical landmarks for endosonography of the larynx.

Methods

In an experimental study, a total of 32 normal human larynges were examined endosonographically, and the classical landmarks were correlated to horizontal whole‐organ sections of the scanned specimens.

Results

All laryngeal specimens showed a similar and reproducible sonoanatomy, which could be verified consistently on corresponding histological cross sections. Anatomical structures readily identified included the laryngeal framework, the vocal ligament, the vocal muscle, the ventricular fold, the preepiglottic and paraglottic space, and the epiglottis.

Conclusions

Due to a reproducible sonoanatomy of the larynx, endosonography might be an interesting complementary tool in the diagnostic investigation of laryngeal lesions such as medium‐sized tumors, cysts, laryngoceles, and stenoses. © 2009 Wiley Periodicals, Inc. Head Neck, 2010
Malakoplakia presenting as an enlarging neck mass: Case presentation and review of the literature
Authors: Garg Montey , Eley Karen A. , Bond Stephen E. , Shah Ketan A. , Browning Lisa , Watt‐Smith Stephen R.
Published: July 2009

Background.

Malakoplakia is a very rare chronic inflammatory disorder, first described in 1902. In 75% of cases, the condition affects the genitourinary tract. Five cases of malakoplakia affecting the neck were previously reported in the literature.

Methods and Results.

An 83‐year‐old woman presented with an enlarging mass in the posterior triangle of the neck that was histologically confirmed as malakoplakia. Presenting features are often nonspecific, and the diagnosis is dependent on histological findings. The characteristic microscopic findings are of Michaelis–Gutmann (M‐G) bodies that stain positive with periodic acid–Schiff reagent, von Kossa's reaction for calcium, and Perl's ferrocyanide reaction to ferric iron.

Conclusion.

Although rare, a diagnosis of malakoplakia should be considered in patients with an enlarging mass. This may mimic the presentation of malignancy, particularly in patients in whom erosion through skin occurs, and histological confirmation is advocated. © 2009 Wiley Periodicals, Inc. Head Neck 32:1269–1272, 2010
Detection of cervical metastases with 11C‐tyrosine pet in patients with squamous cell carcinoma of the oral cavity or oropharynx: A comparison with 18F‐FDG PET
Authors: Krabbe Christiaan A. , van der Werff‐Regelink Gerreke , Pruim Jan , van der Wal Jacqueline E. , Roodenburg Jan L. N.
Published: July 2009

Background.

A disadvantage of 2‐[18F]fluoro‐2‐deoxy‐D‐glucose (18F‐FDG) positron emission tomography (PET) in head and neck cancer is that 18F‐FDG uptake is not specific to malignant tissue. To provide an alternative, radiolabeled amino acids such as L‐1‐[11C]‐tyrosine (11C‐TYR), were introduced because these are less avidly metabolized by inflammatory cells.

Methods.

In this prospective study, we compared both 11C‐TYR PET and 18F‐FDG PET performance in detecting cervical metastases in 27 patients with a squamous cell carcinoma (SCC) of oral cavity or oropharynx.

Results.

11C‐TYR PET sensitivity, specificity, and accuracy for detecting nodal metastases were 33%, 100%, and 81%, respectively. With respect to 18F‐FDG PET, these figures were 67%, 97%, and 89%, respectively. Neck metastases not detected by 11C‐TYR PET were camouflaged by high tracer uptake by salivary glands.

Conclusions.

Because of bilateral accumulation of 11C‐TYR in salivary glands, 11C‐TYR PET is not suitable to replace 18F‐FDG PET in staging SCC of oral cavity and oropharynx. © 2009 Wiley Periodicals, Inc. Head Neck, 2010
Absence of Simian virus 40, BK, and JC polyomavirus DNA in squamous cell carcinoma limited to the oral cavity
Authors: Palmieri Annalisa , Carinci Francesco , Martinelli Marcella , Spinelli Giuseppe , Lo Muzio Lorenzo , Rubini Corrado , Scapoli Luca
Published: July 2009

Background.

Head and neck squamous cell carcinomas (SCCs) are among the most aggressive types of cancer. The Simian virus 40 (SV40), which is a polyomavirus known for its oncogenic potential, was found as a contaminant of oral vaccines and has been related to human pleomorphic adenoma in the parotid gland. The aim of this study was to evaluate the presence of SV40 and 2 human polyomaviruses—BK virus (BKV) and JC virus (JCV)—in a large sample of SCCs of the oral cavity.

Methods.

Quantitative real‐time polymerase chain reaction (PCR) was used to evaluate virus load.

Results.

Overall, the prevalence of SV40, BKV, and JCV in oral SCC was negligible. Matched‐pair case‐control analysis indicated that prevalence among the controls did not significantly differ with respect to analyzed cases.

Conclusion.

The results did not indicate a major role for SV40, BKV, and JCV in the etiology of oral SCC. © 2009 Wiley Periodicals, Inc. Head Neck, 2010
Non‐toxic melanoma therapy by a novel tubulin‐binding agent
Authors: Aneja Ritu , Asress Seneshaw , Dhiman Neerupma , Awasthi Anshumali , Rida Padmashree C.G. , Arora Sudarshan K. , Zhou Jun , Glass Jonathan D. , Joshi Harish C.
Published: July 2009

(S)‐3‐((R)‐9‐bromo‐4‐methoxy‐6‐methyl‐5,6,7,8‐tetrahydro‐[1,3]dioxolo[4,5‐g]isoquino‐lin‐5‐yl)‐6,7‐dimethoxyisobenzofuran‐1(3H)‐one (EM011) is a tubulin‐binding agent with significant anticancer activity. Here we show that EM011 modulates microtubule dynamics at concentrations that do not alter the total polymer mass of tubulin. In particular, EM011 decreases the transition frequencies between growth and shortening phases and increases the duration microtubules spend in an idle ‘pause’ state. Using B16LS9 murine melanoma cells, we show that EM011 briefly arrests cell‐cycle progression at the G2/M phase by formation of multiple aster spindles. An aberrant mitotic exit without cytokinesis then occurs, leading to the accumulation of abnormal multinucleated cells prior to apoptosis. Our pharmacokinetic studies conformed to a linear dose‐response relationship upto 150 mg/kg. However, non‐linearity was observed at 300 mg/kg. In a syngeneic murine model of subcutaneous melanoma, better antitumor responses were seen at 150 mg/kg compared to 300 mg/kg of EM011. Unlike currently available chemotherapeutics, EM011 is non‐toxic to normal tissues and most importantly, does not cause any immunosuppression and neurotoxicity. Our data thus warrant a clinical evaluation of EM011 for melanoma therapy.
Synergistic actions of atorvastatin with γ‐tocotrienol and celecoxib against human colon cancer HT29 and HCT116 cells
Authors: Yang Zhihong , Xiao Hang , Jin Huanyu , Koo Phillip T. , Tsang Dorothea J. , Yang Chung S.
Published: July 2009

The synergistic actions of atorvastatin (ATST) with γ‐tocotrienol (γ‐TT) and celecoxib (CXIB) were studied in human colon cancer cell lines HT29 and HCT116. The synergistic inhibition of cell growth by ATST and γ‐TT was demonstrated by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and isobologram analysis. δ‐TT exhibited a similar inhibitory action when combined with ATST. Mevalonate and geranylgeranyl pyrophosphate eliminated most of the growth inhibitory effect of ATST, but only marginally decreased that of γ‐TT; whereas farnesyl pyrophosphate and squalene exhibited little effect on the inhibitory action of ATST and γ‐TT, indicating protein geranylgeranylation, but not farnesylation are involved in the inhibition of colon cancer cell growth. Both mevalonate and squalene restored the cellular cholesterol level that was reduced by ATST treatment, but only mevalonate eliminated the cell growth inhibitory effect, suggesting that the cholesterol level in cells does not play an essential role in inhibiting cancer cell growth. Protein level of HMG‐CoA reductase increased after ATST treatment, and the presence of γ‐TT attenuated the elevated level of HMG‐CoA reductase. ATST also decreased membrane‐bound RhoA, possibly due to a reduced level of protein geranylgeranylation; addition of γ‐TT enhanced this effect. The mediation of HMG‐CoA reductase and RhoA provides a possible mechanism for the synergistic action of ATST and γ‐TT. The triple combination of ATST, γ‐TT and CXIB showed a synergistic inhibition of cancer cell growth in MTT assays. The synergistic action of these three compounds was also illustrated by their induction of G0/G1 phase cell cycle arrest and apoptosis.
Rlip76 transports sunitinib and sorafenib and mediates drug resistance in kidney cancer
Authors: Singhal Sharad S. , Sehrawat Archana , Sahu Mukesh , Singhal Preeti , Vatsyayan Rit , Rao Lelsani Poorna Chandra , Yadav Sushma , Awasthi Sanjay
Published: July 2009

RLIP76 is a stress‐responsive membrane protein implicated in the regulation of multiple cellular signaling pathways. It represents the predominant glutathione‐conjugate (GS‐E) transporter in cells. We have shown that RLIP76 plays a crucial role in defending cancer cells from radiation and chemotherapeutic toxin‐mediated apoptosis, and that its inhibition by antibodies or depletion by siRNA or antisense causes apoptosis in a number of cancer cell types. We demonstrated for the first time that the striking anti‐neoplastic effects with no evident toxicity in terms of either weight loss or metabolic effects are also demonstrable for the antibody, antisense and siRNA in a renal cell xenografts model of Caki‐2 cells (Singhal et al., Cancer Res., 2009, 69: 4244). Present studies were performed to determine if RLIP76 targeting is more broadly applicable in other kidney cancer cell lines, to compare the signaling effects of RLIP76 antisense with kinase inhibitors used in treatment of renal cell carcinoma, and to determine whether kinase inhibitors were substrates for transport by RLIP76. Results of these studies show that sorafenib as well as sunitinib are substrates for transport by RLIP76 thus are competitive inhibitors of GS‐E transport. Furthermore, kinase inhibition in the ERK as well as PI3K pathways by RLIP76 depletion is more profound and consistent and is more widely apparent in a number of renal carcinoma cell lines. These studies offer strong support for our overall hypothesis that RLIP76 is an overarching anti‐apoptosis mechanism that, if inhibited, can be more broadly effective in the treatment of renal cell carcinoma.
Outcome prediction and risk assessment by quantitative pyrosequencing methylation analysis of the SFN gene in advanced stage, high‐risk, neuroblastic tumor patients
Authors: Banelli Barbara , Bonassi Stefano , Casciano Ida , Mazzocco Katia , Di Vinci Angela , Scaruffi Paola , Brigati Claudio , Allemanni Giorgio , Borzì Luana , Tonini Gian Paolo , Romani Massimo
Published: July 2009

The aim of our study was to identify threshold levels of DNA methylation predictive of the outcome to better define the risk group of stage 4 neuroblastic tumor patients. Quantitative pyrosequencing analysis was applied to a training set of 50 stage 4, high risk patients and to a validation cohort of 72 consecutive patients. Stage 4 patients at lower risk and ganglioneuroma patients were included as control groups. Predictive thresholds of methylation were identified by ROC curve analysis. The prognostic end points of the study were the overall and progression‐free survival at 60 months. Data were analyzed with the Cox proportional hazard model. In a multivariate model the methylation threshold identified for the SFN gene (14.3.3σ) distinguished the patients presenting favorable outcome from those with progressing disease, independently from all known predictors (Training set: Overall Survival HR 8.53, p = 0.001; Validation set: HR 4.07, p = 0.008). The level of methylation in the tumors of high‐risk patients surviving more than 60 months was comparable to that of tumors derived from lower risk patients and to that of benign ganglioneuroma. Methylation above the threshold level was associated with reduced SFN expression in comparison with samples below the threshold. Quantitative methylation is a promising tool to predict survival in neuroblastic tumor patients. Our results lead to the hypothesis that a subset of patients considered at high risk—but displaying low levels of methylation—could be assigned at a lower risk group.
Role of ERCC1 promoter hypermethylation in drug resistance to cisplatin in human gliomas
Authors: Chen Hua‐Yun , Shao Cui‐Jie , Chen Fu‐Rong , Kwan Aij‐Lie , Chen Zhong‐Ping
Published: July 2009

Overexpression of ERCC1 mRNA is associated with drug resistance to cisplatin in human gliomas, but the role of the ERCC1 promoter in drug resistance has not been demonstrated. We have used sodium bisulfite sequencing to compare ERCC1 promoter methylation patterns in cisplatin‐sensitive and cisplatin‐resistant glioma cells. The levels of ERCC1 DNA methylation, mRNA and protein in 32 human glioma samples were examined by methylation specific PCR, real‐time RT‐PCR and immunohistochemistry, respectively. Meanwhile, cisplatin sensitivities to these human glioma samples were tested by histoculture drug response assay. Hypermethylation was observed in the upstream 5Kb region of the ERCC1 promoter of cisplatin‐sensitive glioma cell lines. ERCC1 DNA methylation levels were highly variable in 32 human glioma samples ranging from 0.1 to 0.87, which have shown significant difference between cisplatin‐sensitive samples and cisplatin‐resistant samples (p < 0.05). The relative expression levels of ERCC1 mRNA in 32 glioma samples were also variable from 0.01 to 5.71. No detectable or low expression of ERCC1 protein was shown in 7 glioma samples. ERCC1 promoter methylation was inversely correlated to mRNA expression (r = −0.903 p = 0.001) as well as protein expression (r = −0.884 p = 0.001). Moreover, ERCC1 mRNA expression was significantly associated with protein levels (r = 0.840 p = 0.001). In summary, the aberrant CpG island methylation in ERCC1 promoter region exists in human glioma cell lines as well as clinical glioma samples. ERCC1 DNA methylation could regulate the expression of downstream mRNA and protein, and was associated with cisplatin chemosensitivity.
Energy dispersive X‐ray analysis of titanium dioxide nanoparticle distribution after intravenous and subcutaneous injection in mice
Authors: Patri Anil , Umbreit Thomas , Zheng J. , Nagashima K. , Goering Peter , Francke‐Carroll Sabine , Gordon Edward , Weaver James , Miller Terry , Sadrieh Nakissa , McNeil Scott , Stratmeyer Mel
Published: July 2009

In an effort to understand the disposition and toxicokinetics of nanoscale materials, we used EDS (energy dispersive X‐ray spectroscopy) to detect and map the distribution of titanium dioxide (TiO2) in tissue sections from mice following either subcutaneous (s.c.) or intravenous (i.v.) injection. TiO2 nanoparticles were administered at a dose of 560 mg/kg (i.v.) or 5600 mg/kg (s.c.) to Balb/c female mice on two consecutive days. Tissues (liver, kidney, lung, heart, spleen, and brain) were examined by light microscopy, TEM (transmission electron microscopy), SEM (scanning electron microscopy), and EDS following necropsy one day after treatment. Particle agglomerates were detected by light microsopy in all tissues examined, EDS microanalysis was used to confirm that these tissues contained elemental titanium and oxygen. The TEM micrographs and EDS spectra of the aggregates were compared with in vitro measurements of TiO2 nanoparticle injection solution (i.e., in water). The nanoparticles were also characterized using dynamic light scattering in water, 10 mM NaCl, and phosphate buffered saline (PBS). In low ionic strength solvents (water and 10 mM NaCl), the TiO2 particles had average hydrodynamic diameters ranging from 114–122 nm. In PBS, however, the average diameter increases to 1–2 μm, likely due to aggregation analogous to that observed in tissue by TEM and EDS. This investigation demonstrates the suitability of energy dispersive X‐ray spectroscopy (EDS) for detection of nanoparticle aggregates in tissues and shows that disposition of TiO2 nanoparticles depends on the route of administration (i.v. or s.c.). Published in 2009 by John Wiley and Sons, Ltd.
Analyzing cell fate control by cytokines through continuous single cell biochemistry
Authors: Rieger Michael A. , Schroeder Timm
Published: July 2009

Cytokines are important regulators of cell fates with high clinical and commercial relevance. However, despite decades of intense academic and industrial research, it proved surprisingly difficult to describe the biological functions of cytokines in a precise and comprehensive manner. The exact analysis of cytokine biology is complicated by the fact that individual cytokines control many different cell fates and activate a multitude of intracellular signaling pathways. Moreover, although activating different molecular programs, different cytokines can be redundant in their biological effects. In addition, cytokines with different biological effects can activate overlapping signaling pathways. This prospect article will outline the necessity of continuous single cell biochemistry to unravel the biological functions of molecular cytokine signaling. It focuses on potentials and limitations of recent technical developments in fluorescent time‐lapse imaging and single cell tracking allowing constant long‐term observation of molecules and behavior of single cells. J. Cell. Biochem. 108: 343–352, 2009. © 2009 Wiley‐Liss, Inc.
Distribution of TGF‐β isoforms and signaling intermediates in corneal fibrotic wound repair
Authors: Huh Man‐IL , Kim Yeoun‐Hee , Park Jong‐Hyuck , Bae Sung‐Won , Kim Min‐Hee , Chang Yongmin , Kim Song‐Ja , Lee Sun‐Ryung , Lee Young‐Sup , Jin Eun‐Jung , Sonn Jong‐Kyung , Kang Shin‐Sung , Jung Jae‐Chang
Published: July 2009

In this study, temporal and spatial distribution of three TGF‐β isoforms and their downstream signaling pathways including pSmad2 and p38MAPK were examined during fibrotic wound repair. In normal chick corneas, TGF‐β1, ‐2, and ‐3 were weakly detected in Bowman's layer (BL). In healing corneas, TGF‐β1 was primarily deposited in the fibrin clot and the unwounded BL. TGF‐β2 was highly expressed in healing epithelial and endothelial cells, and numerous active fibroblasts/myofibroblasts. TGF‐β3 was mainly detected in the unwound region of basal epithelial cells. α‐Smooth muscle actin (α‐SMA) was initially appeared in the posterior region of repairing stroma at day 3, and was detected in the entire healing stroma by day 7. Notably, α‐SMA was absent in the central region of healing stroma by day 14, and its staining pattern was similar to those of TGF‐β2 and p38MAPK. By contrast, pSmad2 was mainly detected in the fibroblasts. In normal cornea, laminin was mainly detected in both epithelial basement membrane (BM) and Descemet's membrane (DM). By contrast to reconstitution of the BM in the wound region, the DM was not repaired although endothelial layer was regenerated, indicating that high levels of TGF‐β2 were released into the posterior region of healing stroma on day 14. High levels of α‐SMA staining, shown in cultured repair stromal cells from healing corneas on day 14 and in TGF‐β2 treated normal stromal cells, were significantly reduced by p38MAPK inhibition. Collectively, this study suggests that TGF‐β2‐mediated myofibroblast transformation is mediated, at least partly, by the p38MAPK pathway in vivo. J. Cell. Biochem. 108: 476–488, 2009. © 2009 Wiley‐Liss, Inc.
Glucosamine inhibits IL‐1β‐mediated IL‐8 production in prostate cancer cells by MAPK attenuation
Authors: Tsai Cheng‐Yen , Lee Tzong‐Shyuan , Kou Yu Ru , Wu Yuh‐Lin
Published: July 2009

Inflammation is a complex process involving cytokine production to regulate host defense cascades. In contrast to the therapeutic significance of acute inflammation, a pathogenic impact of chronic inflammation on cancer development has been proposed. Upregulation of inflammatory cytokines, such as IL‐1β and IL‐8, has been noted in prostate cancer patients and IL‐8 has been shown to promote prostate cancer cell proliferation and migration; however, it is not clear whether IL‐1β regulates IL‐8 expression in prostate cancer cells. Glucosamine is widely regarded as an anti‐inflammatory agent and thus we hypothesized that if IL‐1β activated IL‐8 production in prostate cancer cells, then glucosamine ought to blunt such an effect. Three prostate cancer cell lines, DU‐145, PC‐3, and LNCaP, were used to evaluate the effects of IL‐1β and glucosamine on IL‐8 expression using ELISA and RT‐PCR analyses. IL‐1β elevated IL‐8 mRNA expression and subsequent IL‐8 secretion. Glucosamine significantly inhibited IL‐1β‐induced IL‐8 secretion. IL‐8 appeared to induce LNCaP cell proliferation by MTT assay; involvement of IL‐8 in IL‐1β‐dependent PC‐3 cell migration was demonstrated by wound‐healing and transwell migration assays. Inhibitors of MAPKs and NFκB were used to pinpoint MAPKs but not NFκB being involved in IL‐1β‐mediated IL‐8 production. IL‐1β‐provoked phosphorylation of all MAPKs was notably suppressed by glucosamine. We suggest that IL‐1β can activate the MAPK pathways resulting in an induction of IL‐8 production, which promotes prostate cancer cell proliferation and migration. In this context, glucosamine appears to inhibit IL‐1β‐mediated activation of MAPKs and therefore reduces IL‐8 production; this, in turn, attenuates cell proliferation/migration. J. Cell. Biochem. 108: 489–498, 2009. © 2009 Wiley‐Liss, Inc.
Involvement of STIM1 in the proteinase‐activated receptor 1‐mediated Ca2+influx in vascular endothelial cells
Authors: Hirano Katsuya , Hirano Mayumi , Hanada Akiko
Published: July 2009

Thrombin increases the cytosolic Ca2+concentrations and induces NO production by activating proteinase‐activated receptor 1 (PAR1) in vascular endothelial cells. The store‐operated Ca2+influx is a major Ca2+influx pathway in non‐excitable cells including endothelial cells and it has been reported to play a role in the thrombin‐induced Ca2+signaling in endothelial cells. Recent studies have identified stromal interaction molecule 1 (STIM1) to function as a sensor of the store site Ca2+content, thereby regulating the store‐operated Ca2+influx. However, the functional role of STIM1 in the thrombin‐induced Ca2+influx and NO production in endothelial cells still remains to be elucidated. Fura‐2 and diaminorhodamine‐4M fluorometry was utilized to evaluate the thrombin‐induced changes in cytosolic Ca2+concentrations and NO production, respectively, in porcine aortic endothelial cells transfected with small interfering RNA (siRNA) targeted to STIM1. STIM1‐targeted siRNA suppressed the STIM1 expression and the thapsigargin‐induced Ca2+influx. The degree of suppression of the STIM1 expression correlated well to the degree of suppression of the Ca2+influx. The knockdown of STIM1 was associated with a substantial inhibition of the Ca2+influx and a partial reduction of the NO production induced by thrombin. The thrombin‐induced Ca2+influx exhibited the similar sensitivity toward the Ca2+influx inhibitors to that seen with the thapsigargin‐induced Ca2+influx. The present study provides the first evidence that STIM1 plays a critical role in the PAR1‐mediated Ca2+influx and Ca2+‐dependent component of the NO production in endothelial cells. J. Cell. Biochem. 108: 499–507, 2009. © 2009 Wiley‐Liss, Inc.
Negative regulation of endogenous protein kinase Cα on the dynamic change of carbachol‐induced intracellular calcium response in different melanoma cells
Authors: Wang Huan , Yu Yao‐Qing , Liao Wen‐Jun , Wang Zong‐Ren , Lv Ya‐Jie , Zhang Yan‐Guo , Gao Tian‐Wen
Published: July 2009

Regulations of intracellular protein kinase C (PKC) on carbachol (CCh)‐induced intracellular calcium ([Ca2+]i) responses were investigated in different stages of melanoma cells. We found that CCh (1 mM) significantly increased [Ca2+]i with 6‐, 4‐, 4‐, and 25‐folds intensities in WM793B, 451Lu, SK‐MEL‐5, and A2058 melanoma cells, respectively. Pretreatment of phorbol 12, 13‐dibutyrate (PDBu, 2 µM), an activator of intracellular PKC, significantly suppressed CCh‐induced peak reactions in WM793B, SK‐MEL‐5, and A2058 cells. RT‐PCR data showed that mRNA levels of PKCα were 12‐, 4‐, 6‐, and 0.9‐folds higher in above four melanoma cells. Short interfering RNA (siRNA) targeting to PKCα in WM793B cells enhanced CCh‐induced peak calcium reactions. Present data indicated that CCh‐induced [Ca2+]i responses were dynamically changed in different stages of melanoma progression. Moreover, intracellular PKCα activated by exogenous agonist and expressed through endogenous gene transcription negatively regulated CCh‐induced calcium responses. The functional analysis on the relationship between CCh‐induced calcium response and endogenous PKCα expression might be helpful to predict the development of melanoma. J. Cell. Physiol. 221: 276–282, 2009. © 2009 Wiley‐Liss, Inc.
Regulation of HIV‐1 transcription at 3% versus 21% oxygen concentration
Authors: Charles Sharroya , Ammosova Tatyana , Cardenas Jessica , Foster Altreisha , Rotimi Jamie , Jerebtsova Marina , Ayodeji Abisola A. , Niu Xiaomei , Ray Patricio E. , Gordeuk Victor R. , Kashanchi Fatah , Nekhai Sergei
Published: July 2009

HIV transcription is induced by the HIV‐1 Tat protein, in concert with cellular co‐factors including CDK9, CDK2, NF‐κB, and others. The cells of most of the body's organs are exposed to ∼3–6% oxygen, but most in vitro studies of HIV replication are conducted at 21% oxygen. We hypothesized that activities of host cell factors involved in HIV‐1 replication may differ at 3% versus 21% O2, and that such differences may affect HIV‐1 replication. Here we show that Tat‐induced HIV‐1 transcription was reduced at 3% O2 compared to 21% O2. HIV‐1 replication was also reduced in acutely or chronically infected cells cultured at 3% O2 compared to 21% O2. This reduction was not due the decreased cell growth or increased cellular toxicity and also not due to the induction of hypoxic response. At 3% O2, the activity of CDK9/cyclin T1 was inhibited and Sp1 activity was reduced, whereas the activity of other host cell factors such as CDK2 or NF‐κB was not affected. CDK9‐specific inhibitor ARC was much less efficient at 3% compared to 21% O2 and also expression of CDK9/cyclin T1‐dependent IκB inhibitor α was repressed. Our results suggest that lower HIV‐1 transcription at 3% O2 compared to 21% O2 may be mediated by lower activity of CDK9/cyclin T1 and Sp1 at 3% O2 and that additional host cell factors such as CDK2 and NF‐κB might be major regulators of HIV‐1 transcription at low O2 concentrations. J. Cell. Physiol. 221: 469–479, 2009. © 2009 Wiley‐Liss, Inc.
293 cells over‐expressing human ADI1 and CD81 are permissive for serum‐derived hepatitis C virus infection
Authors: Cheng Ju‐Chien , Yeh Yung‐Ju , Pai Li‐Mei , Chang Ming‐Ling , Yeh Chau‐Ting
Published: July 2009

Human aci‐reductone dioxygenase 1 (ADI1) is a member of the Cupin superfamily. It binds to and inhibits the activities of membrane‐type 1 matrix metalloproteinase, a protein known to interact with the tight junction protein, claudin‐1. Previously, a variant protein, named submergence‐induced protein‐like factor (Sip‐L), consisting of ADI1 amino acids 64–179, was found to support hepatitis C virus (HCV) infection and replication in 293 cells. In the present study, it was discovered that over‐expression of human ADI1 in 293 cells (293‐ADI1 cells) also supported HCV infection and replication. Using serum‐derived HCV as an infectious source, enhanced cell uptake of HCV to a Northern blot detectable level was found in 293 cells over‐expressing both CD81 and ADI1 (293‐ADI1‐CD81 cells). The enhanced cell entry was confirmed by the use of the vesicular stomatitis virus‐based HCV pseudotype particles. However, transfection of HCV replicon RNA by electroporation into naïve 293 and 293‐ADI1 cells revealed no difference in replication efficiency. Using the infectious J6/JFH chimera as an infectious source, the infectivity was compared between 293‐ADI1‐CD81 and Huh‐7.5 cells. More infection foci were formed in the 293‐ADI1‐CD81 cells in the first round of infection. In conclusion, human ADI1 over‐expression in 293 cells enhances cell entry but not replication of HCV. 293‐ADI1‐CD81 cells are permissive for serum‐derived HCV infection. J. Med. Virol. 81:1560–1568, 2009. © 2009 Wiley‐Liss, Inc.