bluuebunny/crossref_metadata_2025 huggingface dataset:
Healthier lipid combination oil‐in‐water emulsions prepared with various protein systems: an approach for development of functional meat products
Authors: Delgado‐Pando Gonzalo , Cofrades Susana , Ruiz‐Capillas Claudia , Teresa Solas María , Jiménez‐Colmenero Francisco
Published: April 2010

Five protein‐stabilized oil‐in‐water emulsions were prepared using sodium caseinate (O/SC), soy protein isolate (O/SPI), sodium caseinate and microbial transglutaminase (O/SC + MTG), sodium caseinate, microbial transglutaminase and meat slurry (O/SC + MTG + MS) and SPI, sodium caseinate and microbial transglutaminase (O/IPS + SC + MTG); their composition (proximate analysis and fatty acid profile) and physicochemical characteristics were examined. The lipid phase was a combination of healthy fatty acids from olive, linseed and fish oils, containing low proportions (15%) of saturated fatty acids (SFA) and high proportions of monounsaturated fatty acids (MUFA, 47%) and polyunsaturated fatty acids (PUFA, 36%), with a PUFA/SFA ratio >2, and a n‐6/n‐3 PUFA ratio of 0.4. All the oil‐in‐water emulsions showed high thermal and creamy stability. Results of penetration test and dynamic rheological properties showed la existencia de different types of oil‐in‐water emulsion structures according to stabilizing system of emulsion. Those structures ranged from concentrate solution‐like (stabilized only with SC) (gel strength 0.06 mJ) to gel‐like (samples containing MTG) behaviours (gel strength ranged between 3.4 and 6.2 mJ). Morphological differences in the organization of the network structure were observed (by scanning electron microscopy) as functions of the protein system used to stabilize the oil‐in‐water emulsions.
Improvement of mono and diacylglycerol production via enzymatic glycerolysis in tert‐butanol system
Authors: Krüger Roberta L. , Valério Alexsandra , Balen Manuela , Ninow Jorge Luiz , Vladimir Oliveira J. , de Oliveira Débora , Corazza Marcos Lúcio
Published: April 2010

In this work we report experimental data regarding the glycerolysis of olive oil using Novozym 435 in tert‐butanol organic system aiming at the production of monoacylglycerols (MAG) and diacylglycerols (DAG). Experiments were performed in batch mode, recording the reaction kinetics and evaluating the effects of temperature, enzyme concentration, tert‐butanol:oil/glycerol volume ratio and using solvent to substrates ratio of 1:1 and 5:1 v/v. Experimental results showed that lipase‐catalyzed glycerolysis in tert‐butanol might be a potential route for the production of high contents of MAG and DAG. The results also showed that it is possible to maximize the production of MAG and/or DAG, depending on the glycerol to oil molar ratio employed in the reactional system. Higher contents of MAG (53 wt%) and DAG (50 wt%) were achieved using glycerol to oil molar ratio of 3:1/6:1 and 0.5:1.5, respectively, both in 8 h of reaction at 70°C, 600 rpm and enzyme concentration of 10 wt%.
Improvement of fuel properties of cottonseed oil methyl esters with commercial additives
Authors: Joshi Hem , Moser Bryan R. , Shah Shailesh N. , Mandalika Anurag , Walker Terry
Published: April 2010

The low temperature operability and oxidative stability of cottonseed oil methyl esters (CSME) were improved with four anti‐gel additives as well as one antioxidant additive, gossypol. Low temperature operability and oxidative stability of CSME was determined by cloud point (CP), pour point (PP), cold filter plugging point (CFPP), and oxidative stability index (OSI). The most significant reductions in CP, PP, and CFPP in all cases were obtained with Technol®, with the average reduction in temperature found to be 3.9 °C. Gunk®, Heet®, and Howe's® were progressively less effective, as indicated by average reductions in temperature of 3.4, 3.0, and 2.8 °C, respectively. In all cases, the magnitude of CFPP reduction was greater than for PP and especially CP. Addition of gossypol, a polyphenolic aldehyde, resulted in linear improvement in OSI (R2 = 0.9804). The OSI of CSME increased from 5.0 to 8.3 h with gossypol at a concentration of 1000 ppm.
Rhamnolipid production by Burkholderia plantarii DSM 9509T
Authors: Hörmann Barbara , Müller Markus M. , Syldatk Christoph , Hausmann Rudolf
Published: April 2010

The Burkholderia plantarii type strain DSM 9509T was shown to produce extracellular rhamnolipids that clearly differ in structure and properties from the Pseudomonas aeruginosa rhamnolipids. The type strain has not been previously described as rhamnolipid producer. B. plantarii DSM 9509T was cultivated in a parallel bioreactor system and produced up to 45.74 mg/L of rhamnolipids when grown in glucose supplemented rich medium. The structure of the main rhamnolipid Rha2‐C14‐C14 (L‐rhamnopyranosyl‐L‐rhamnopyranosyl‐β‐hydroxytetradecanoyl‐β‐hydroxytetradecanoate) was verified by MS/MS. The purified rhamnolipid Rha2‐C14‐C14 lowered the equilibrium surface tension of distilled water to 29.4 mN/m. The CMC was between 15 and 20 mg/L. In this study, growth and product formation were investigated under biotechnological aspects.

Practical applications: B. plantarii DSM 9509T produces a rhamnolipid that differs in structure from P. aeruginosa rhamnolipids and exhibits excellent surfactant properties. An essential advantage in comparison to P. aeruginosa is that B. plantarii DSM 9509T is a Biosafety Level 1 microorganism and not pathogenic in humans. A biotechnological production of the Burkholderia rhamnolipids could lead to their application in detergents, pharmaceuticals, and other industries providing new products on the biosurfactant market.
Disposable Graphite Foil Based Electrodes and Their Application in Pharmaceutical Analysis
Authors: Oliveira André G. , Munoz Rodrigo Alejandro A. , Angnes Lúcio
Published: April 2010

This paper explores a new source of graphite for working electrodes, which presents advantages such as low electrical resistance, good flexibility, favorable mechanical performance, versatility to design electrodes in almost any size and very low cost. The new electrodes were investigated in batch electrochemical cells as associated with flow injection analysis systems. Cyclic voltammetry, stripping voltammetry, and amperometry associated with flow injection analysis techniques were applied for the determination of ascorbic acid, zinc and paracetamol in pharmaceutical formulations, respectively. Well‐established analytical methods were applied for comparison purposes. The results herein demonstrate the potential of graphite foils as working electrodes in different electroanalytical methods, offering the possibility of producing disposable sensors for routine applications.
New Determination Scheme of p‐Aminophenol by MnO2 Modified Electrode Coupled with Flow Injection Analysis
Authors: Lin Meng‐Shan , Jan Bor‐Iuan , Chen Pei‐Yen , Cheng Wen‐Chuan , Chen Chi‐Hao
Published: April 2010

New determination scheme of p‐aminophenol by using MnO2 as a preoxidant is demonstrated in this work. In the flow injection system, the p‐aminophenol is oxidized to quinoneimine by MnO2 at up‐stream, which can be detected at a suitable reductive potential. After optimization, the linear range of PAP is started from 1 μM to 30 μM (R2=0.999), the estimated detection limit (S/N=3) is 0.28 μM. Two real samples are studied and excellent recoveries are achieved by using standard addition method.
Adsorptive Stripping Voltammetry of Nickel at an In Situ Plated Bismuth Film Electrode
Authors: Korolczuk Mieczyslaw , Rutyna Iwona , Tyszczuk Katarzyna
Published: April 2010

For the first time an in situ plated bismuth film electrode is applied to adsorptive stripping voltammetry at mild alkaline conditions, to pH of 10. As an example of the application of such an electrode a procedure for trace Ni(II) determination by adsorptive stripping voltammetry is elaborated. For stabilization of Bi(III), tartrate as a strong complexing agent for Bi(III) was added to an ammonia buffer used as a supporting electrolyte. The appropriate concentration of Bi(III) was added to the electrolyte in the form of its complex with tartrate. The calibration graph for Ni(II) for an accumulation time of 120 s is linear from 5×10−9 to 5×10−8 mol L−1. The proposed procedure was applied for Ni(II) determination in certified water reference material.
Voltammetric Study of Aminopurines on Pencil Graphite Electrode in the Presence of Copper Ions
Authors: Aladag Nilay , Trnkova Libuse , Kourilova Alena , Ozsoz Mehmet , Jelen Frantisek
Published: April 2010

Electrochemical oxidations of aminopurines (adenine, 2‐aminopurine, 2,6‐diaminopurine) and their complexes with Cu(I) on a pencil graphite electrode were investigated by means of linear sweep voltammetry (LSV) and elimination voltammetry with linear scan (EVLS). The anodic process of the Cu(I)‐aminopurine complex, corresponding to the oxidation of Cu(I) to Cu(II), takes place in the potential range between 0.4 and 0.5 V (vs. Ag/AgCl/3 M KCl). At more positive potentials the aminopurines provide voltammetric peaks resulting from the oxidation of the purine ring. The stability of the accumulated complex layer was investigated by the adsorptive transfer stripping technique.
Ecogeomorphic state variables and phase‐space construction for quantifying the evolution of vegetated aeolian landscapes
Authors: Baas Andreas C. W. , Nield Joanna M.
Published: April 2010

Cellular automaton modelling for the simulation of dune field formation and evolution has developed progressively in aeolian geomorphology in the last decade or so. A model that incorporates the effects of vegetation and its interactions with geomorphic landscape development – the Discrete Ecogeomorphic Aeolian Landscapes (DECAL) model – can replicate a number of important visual and qualitative aspects of the complex evolution of aeolian dune landscapes under the influence of vegetation dynamics in coastal environments. A key challenge in this research area is the analysis and comparison of both simulated and real‐world vegetated dune landscapes using objective and quantifiable principles. This study presents a methodological framework or protocol for numerically quantifying various ecogeomorphic attributes, using a suite of mathematically defined landscape metrics, to provide a rigorous and statistical evaluation of vegetated dune field evolution. Within this framework the model parameter space can be systematically explored and simulation outcomes can be methodically compared against real‐world landscapes. Based on a simplified scenario of parabolic dunes developing out of blow‐outs the resulting dune field realizations are investigated as a function of variable growth vigour of two simulated vegetation types (pioneer grass and successional woody shrub) by establishing a typological phase‐diagram of different landscape classes. The set of simulation outcomes furthermore defines a higher‐dimensional phase‐space, whose axes or dimensions can be interpreted by analysing how individual ecogeomorphic landscape metrics, or state variables, contribute to the data distribution. Principal component analysis can reduce this to a visual three‐dimensional (3D) phase‐space where landscape evolution can be plotted as time‐trajectories and where we can investigate the effects of changing environmental conditions partway through a simulation scenario. The use of landscape state variables and the construction of a 3D phase‐space presented here may provide a general template for quantifying many other eco‐geomorphic systems on the Earth's surface. Copyright © 2010 John Wiley & Sons, Ltd.
A review of Late Cretaceous fossil birds from Hungary
Authors: Dyke Gareth J. , Ősi Attila
Published: April 2010

We review the previously described Late Cretaceous (Santonian) bird remains from the Csehbánya Formation in the Bakony Mountains of Hungary, augmenting initial work by Ősi (2008), and add a number of newly collected fossils. All together, the eight fossil specimens so far collected from this site are important to our understanding of avian evolution because they document a large range of taxon body sizes from at least one major lineage (Enantiornithes) and come from a critically undersampled time period in the Cretaceous. Globally, very little fossil bird material has been collected from the middle stages of the Late Cretaceous, the Coniacian and Santonian; most known taxa are either Early Cretaceous (ca. 120 Ma) in age or are from the terminal Campanian and Maastrichtian (ca. 70–65 Ma). Indeed, one of the Csehbánya Formation fossil birds is recognized as a new taxon of large enantiornithine, an avisaurid apparently similar in its largely unfused foot morphology to the Argentine Soroavisaurus and to the North American Avisaurus. The Central European records reviewed in this paper highlight the wide distribution of some Late Cretaceous fossil birds, particularly avisaurid enantiornithines, and lead us to a brief discussion of avian biogeography at the end of the Mesozoic. Copyright © 2010 John Wiley & Sons, Ltd.
There is more to mind reading than having theory of mind concepts: new directions in theory of mind research
Authors: Samson Dana , Apperly Ian A.
Published: April 2010

For more than 30 years, researchers have focused on the important transition that children undergo between the ages of 3 and 5, when they start to solve mind‐reading problems that require reasoning about complex mental states, such as beliefs. The main question for debate has been whether, during that transition, children acquire new concepts about how the mind works (i.e. a more sophisticated ‘theory of mind’) or whether their more general cognitive abilities improve and help them deal with the general task demands. Recently, researchers have started to explore mind‐reading abilities in individuals outside of the classic 3–5 age span, showing early theory of mind abilities in ever‐younger children and infants, but also far from flawless performance in adults. In this article, we show how the results of these two new lines of research converge on the idea that there is more to mind reading than having theory of mind concepts: there are various processes required to efficiently implement theory of mind concepts in our reasoning, and there may be, in fact, multiple mind‐reading routes available. We then highlight the emergent new directions for future research. Copyright © 2010 John Wiley & Sons, Ltd.
Human papillomavirus type distribution in 30,848 invasive cervical cancers worldwide: Variation by geographical region, histological type and year of publication
Authors: Li Ni , Franceschi Silvia , Howell‐Jones Rebecca , Snijders Peter J.F. , Clifford Gary M.
Published: April 2010

Pooled data on human papillomavirus (HPV) type distribution in invasive cervical cancer (ICC) can help to predict the potential impact of HPV type‐specific vaccines and screening tests, and to understand the carcinogenicity of HPV types. We performed a meta‐analysis of HPV type‐specific prevalence data published from 1990 to 2010, including a total of 243 studies and 30,848 ICC. The proportion of ICC associated with HPV16 and/or 18 (HPV16/18) was between 70 and 76% in all world regions except Asia. In Western/Central Asia, 82% of ICC was HPV16/18‐associated compared to only 68% in Eastern Asia. The 12 most common HPV types identified, in order of decreasing prevalence, were HPV16 (57%), 18 (16%), 58, 33, 45, 31, 52, 35, 59, 39, 51 and 56. The prevalence of other types, phylogenetically related to those above, ranged from <0.1% for HPV85 to 0.6% for HPV68. Overall HPV prevalence increased significantly from 85.9% in studies published from 1990 to 1999 to 92.9% in studies published from 2006 to 2010. Prevalence increases were large for multiple infections (from 4.0 to 15.7%) and for HPV16 (from 51.8 to 60.0%, including HPV16 alone or in multiple infections). Smaller but significant increases in prevalence were also seen for HPV39, 53 and 58. A large amount of recently published data has improved the understanding of the contribution of a broad range of HPV types to ICC in different world regions. However, estimating the fraction of ICC attributable to different types is increasingly complicated by the detection of multiple HPV infections in ICC.
PD‐1 and PD‐L1 upregulation promotes CD8+ T‐cell apoptosis and postoperative recurrence in hepatocellular carcinoma patients
Authors: Shi Feng , Shi Ming , Zeng Zhen , Qi Rui‐Zhao , Liu Zhen‐Wen , Zhang Ji‐Yuan , Yang Yong‐Ping , Tien Po , Wang Fu‐Sheng
Published: April 2010

Programmed death 1 (PD‐1) and its ligand (PD‐L1) play pivotal roles in regulating host immune responses. However, the inhibitory effects of this pathway on the function of cytotoxic CD8+ T lymphocytes, the main effector cells in hepatocellular carcinoma (HCC) patients, are not well defined. In this study, we characterized circulating and intratumor PD‐1/PD‐L1 expression and analyzed their association with disease progression in a cohort of hepatitis B virus‐infected patients, including 56 with HCC, 20 with liver cirrhosis (LC) and 20 healthy controls (HC). The frequency of circulating PD‐1+CD8+ T cells increased with disease progression from LC to HCC patients versus HC. Furthermore, tumor‐infiltrating effector CD8+ T cells showed a drastic increase in PD‐1 expression. These increases in circulating and intratumor PD‐1+CD8+ T cells could predict poorer disease progression and postoperative recurrence. Immunohistochemical staining showed that PD‐L1 expressing hepatoma cells and apoptotic infiltrating CD8+ T cells were both enriched in tumor sections. In vitro, CD8+ T cells induced PD‐L1 expression on hepatoma cells in an IFN‐γ–dependent manner, which in turn promoted CD8+ T cells apoptosis, and blocking PD‐L1 reversed this effect. Therefore, this study extends our knowledge of the role of the PD‐1/PD‐L1 pathway in tumor evasion and provides evidence for a new therapeutic target in HCC patients.
Frequent changes in subtelomeric DNA methylation patterns and its relevance to telomere regulation during human hepatocarcinogenesis
Authors: Oh Bong‐Kyeong , Um Tae‐Hee , Choi Gi Hong , Park Young Nyun
Published: April 2010

Subtelomeric chromatin modifications are important regulators of telomere length. We examined the subtelomeric DNA methylation status of 7q, 8q, 17q, 18p, 21q and XpYp in 32 pairs of hepatocellular carcinomas (HCCs) and their adjacent non‐HCCs via methylation‐specific PCR (quantified as methylation ratio). In addition, 10q was subjected to bisulfite‐genomic‐sequencing. Telomere length was determined by Southern hybridization. In all cases, the relationship between methylation ratio and telomere length was determined. High levels of methylation ratio were found on chromosomes 7q, 18p and XpYp, whereas 8q 17q and 21q were less methylated in both HCCs and non‐HCCs. Compared to non‐HCCs, HCCs exhibited a higher methylation ratio on 18p and 21q, and a wider distribution of methylation ratio on 7q, 21q and 10q (p < 0.05). The methylation ratio of 18p and of 21q was negatively and positively correlated with telomere length of HCCs, respectively (p < 0.05). We evaluated changes in methylation pattern between non‐HCCs and HCCs. Out of 185 sites, hypermethylation changes from non‐HCC to HCC were found at 47 sites and hypomethylation changes at 31 sites. Changes in methylation pattern were observed at three to four sites among six chromosomal sites in 15 patients (47%). There was a tendency toward hypomethylation changes at 7q (p = 0.013) and hypermethylation changes at 21q (p = 0.057) when telomere lengthened from non‐HCCs to HCCs. In summary, subtelomeric methylation patterns dynamically changed during hepatocarcinogenesis. Subtelomeric methylation at certain regions was related to telomere lengthening or shortening, suggesting an association between subtelomeric chromatin structure and telomere length regulation in human hepatocarcinogenesis.
An ISCOM vaccine combined with a TLR9 agonist breaks immune evasion mediated by regulatory T cells in an orthotopic model of pancreatic carcinoma
Authors: Jacobs Collin , Duewell Peter , Heckelsmiller Klaus , Wei Jiwu , Bauernfeind Franz , Ellermeier Jonathan , Kisser Ulrich , Bauer Christian A. , Dauer Marc , Eigler Andreas , Maraskovsky Eugene , Endres Stefan , Schnurr Max
Published: April 2010

Vaccines based on immune stimulatory complexes (ISCOM) induce T‐cell responses against tumor antigen (Ag). However, immune responses are impaired in pancreatic cancer patients. We investigated the efficacy of an ISCOM vaccine in a murine pancreatic carcinoma model. Panc02 cells expressing OVA as a model Ag were induced subcutaneously or orthotopically in the pancreas of C57BL/6 mice. Treatment consisted of an OVA containing ISCOM vaccine, either used alone or in combination with the TLR9 agonist CpG. The ISCOM vaccine effectively induced Ag‐specific CTL capable of killing tumor cells. However, in mice with established tumors CTL induction by the vaccine was inefficient and did not affect tumor growth. Lack of efficacy correlated with increased numbers of Treg. Depletion of Treg with anti‐CD25 mAb restored CTL induction and prolonged survival. Adding low‐dose CpG to the ISCOM vaccine reduced Treg numbers, enhanced CTL responses and induced regression of pancreatic tumors in a CD8+ T cell–dependent manner. Mice cured from the primary tumor mounted a memory T‐cell response against wild‐type Panc02 tumors, indicative of epitope spreading. Combining ISCOM vaccines with TLR agonists is a promising strategy for breaking tumor immune evasion and deserves further evaluation for the treatment of pancreatic carcinoma.
Autoregulation of phospholipase D activity is coupled to selective induction of phospholipase D1 expression to promote invasion of breast cancer cells
Authors: Kang Dong Woo , Park Mi Hee , Lee Young Jun , Kim Hyung Sik , Lindsley Craig W. , Alex Brown H. , Min Do Sik
Published: April 2010

Phospholipase D (PLD) is an important signaling enzyme implicated in the control of many biological processes, including cell proliferation and survival. Despite the importance of the duration and amplitude of PLD signaling in carcinogenesis, mechanisms that regulate PLD expression remain poorly understood. In our study, we define the regulatory components of the machinery that specifies selective PLD1 inductionviasignals propagated through PLD activity. We demonstrate for the first time that establishment of a positive feedback loop that is dependent on enzymatic activity originating from both PLD1 and PLD2 isozymes enhances selective expression of PLD1, but not PLD2. Phosphatidic acid, the product of PLD activity, leads to an increase in the Ras‐ERK/PI3K‐NFκB signaling cascade and enhances binding of NFκB to the PLD1 promoter, consequently inducing selective PLD1 expression in SK‐BR3 breast cancer cells. Moreover, selective PLD inhibitor suppressed epidermal growth factor‐induced matrix metalloproteinase upregulation and invasion by inhibiting PLD1 expression. In conclusion, we propose that autoregulation of PLD activity might be coupled to induction of PLD1 expression, and thereby play a role in carcinogenesis.
Vitamin D receptor rs2228570 polymorphism and invasive ovarian carcinoma risk: Pooled analysis in five studies within the Ovarian Cancer Association Consortium
Authors: Lurie Galina , Wilkens Lynne R. , Thompson Pamela J. , Carney Michael E. , Palmieri Rachel T. , Pharoah Paul D. P. , Song Honglin , Hogdall Estrid , Kjaer Susanne Kruger , DiCioccio Richard A. , McGuire Valerie , Whittemore Alice S. , Gayther Simon A. , Gentry‐Maharaj Aleksandra , Menon Usha , Ramus Susan J. , Goodman Marc T. ,   on behalf of the Ovarian Cancer Association Consortium
Published: April 2010

The association of invasive ovarian carcinoma risk with the functional polymorphism rs2228570 (aka rs10735810; FokI polymorphism) in the vitamin D receptor (VDR) gene was examined in 1820 white non‐Hispanic cases and 3479 controls in a pooled analysis of five population‐based case–control studies within the Ovarian Cancer Association Consortium. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using unconditional logistic regression. Carriers of the rare T allele were at increased risk of ovarian carcinoma compared to women with the CC genotype in all studies combined; each copy of the T allele was associated with a modest 9% increased risk (OR = 1.09; 95% CI: 1.01–1.19; p = 0.04). No significant heterogeneity among studies was observed (p = 0.37) and, after excluding the dataset from the Hawaii study, the risk association for rs2228570 among replication studies was unchanged (OR = 1.09; 95% CI: 1.00–1.19; p = 0.06). A stronger association of rs2228570 with risk was observed among younger women (aged < 50 years versus 50 years or older) (p = 0.04). In all studies combined, the increased risk per copy of the T allele among younger women was 24% (OR = 1.24; 95% CI: 1.04–1.47; p = 0.02). This association remained statistically significant after excluding the Hawaii data (OR = 1.20; 95% CI: 1.01–1.43; p = 0.04). No heterogeneity of the association was observed by stage (p = 0.46), tumor histology (p = 0.98), or time between diagnosis and interview (p = 0.94). This pooled analysis provides further evidence that the VDR rs2228570 polymorphism might influence ovarian cancer susceptibility.
TIMP‐3 promotes apoptosis in nonadherent small cell lung carcinoma cells lacking functional death receptor pathway
Authors: Kallio Janne P. , Hopkins‐Donaldson Sally , Baker Andrew H. , Kähäri Veli‐Matti
Published: April 2010

Tissue inhibitor of metalloproteinases‐3 (TIMP‐3) has previously been identified as a tumor suppressor for adherent malignant and normal cells. TIMP‐3 inhibits adhesion of cells to extracellular matrix and promotes apoptosis through death receptor‐activated, caspase‐8‐mediated pathway. Here, we have studied the effect of adenovirally mediated overexpression of TIMP‐3 on small cell lung cancer (SCLC) cell lines SW2 and N417, which grow in suspension and lack functional caspase‐8. The results show that adenoviral delivery of TIMP‐3 promotes apoptotic cell death in SCLC cells in the absence of caspase‐8 activation. These results suggest TIMP‐3 as a promising therapeutic anticancer protein also in nonadherent malignant cells lacking functional death receptor signaling.
Time‐domain in vivo near infrared fluorescence imaging for evaluation of matriptase as a potential target for the development of novel, inhibitor‐based tumor therapies
Authors: Napp Joanna , Dullin Christian , Müller Friedemann , Uhland Kerstin , Petri Jean Bernhard , van de Locht Andreas , Steinmetzer Torsten , Alves Frauke
Published: April 2010

Proteolytic enzymes expressed on the surface of tumor cells, and thus easily accessible to external interventions, represent useful targets for anticancer and antimetastatic therapies. In our study, we thoroughly evaluated matriptase, a trypsin‐like transmembrane serine protease, as potential target for novel inhibitor‐based tumor therapies. We applied time‐domain near infrared fluorescence (NIRF) imaging to characterize expression and activity of matriptase in vivo in an orthotopic AsPC‐1 pancreatic tumor model in nude mice. We show strong and tumor‐specific binding of intravenously injected Cy5.5 labeled antimatriptase antibody (MT‐Ab*Cy5.5) only to primary AsPC‐1 tumors and their metastases over time within living mice, taking into account fluorescence intensities and fluorescence lifetimes of the applied probes. Specific binding of MT‐Ab*Cy5.5 to tumor sites was confirmed by ex vivo NIRF imaging of tumor tissue, NIRF microscopy and by coregistration of the in vivo acquired NIRF intensity maps to anatomical structures visualized by flat‐panel volume computed tomography (fpVCT) in living mice. Moreover, using an activatable synthetic substrate S*DY‐681 we could clearly demonstrate that matriptase is proteolytically active in vitro as well as in vivo in tumor‐bearing mice, and that application of synthetic active‐site inhibitors having high affinity and selectivity toward matriptase can efficiently inhibit its proteolytic activity for at least 24 hr. We thus successfully applied NIRF imaging in combination with fpVCT to characterize matriptase as a promising molecular target for inhibitor‐based cancer therapies.
PET imaging of tumor angiogenesis in mice with VEGF‐A–targeted 86Y‐CHX‐A″‐DTPA‐bevacizumab
Authors: Nayak Tapan K. , Garmestani Kayhan , Baidoo Kwamena E. , Milenic Diane E. , Brechbiel Martin W.
Published: April 2010

Bevacizumab is a humanized monoclonal antibody that binds to tumor‐secreted vascular endothelial growth factor (VEGF)‐A and inhibits tumor angiogenesis. In 2004, the antibody was approved by the US Food and Drug Administration (FDA) for the treatment of metastatic colorectal carcinoma in combination with chemotherapy. This report describes the preclinical evaluation of a radioimmunoconjugate, 86Y‐CHX‐A″‐DTPA‐bevacizumab, for potential use in Positron Emission Tomography (PET) imaging of VEGF‐A tumor angiogenesis and as a surrogate marker for 90Y‐based radioimmunotherapy. Bevacizumab was conjugated to CHX‐A″‐DTPA and radiolabeled with 86Y. In vivo biodistribution and PET imaging studies were performed on mice bearing VEGF‐A–secreting human colorectal (LS‐174T), human ovarian (SKOV‐3) and VEGF‐A–negative human mesothelioma (MSTO‐211H) xenografts. Biodistribution and PET imaging studies demonstrated highly specific tumor uptake of the radioimmunoconjugate. In mice bearing VEGF‐A–secreting LS‐174T, SKOV‐3 and VEGF‐A–negative MSTO‐211H tumors, the tumor uptake at 3 days postinjection was 13.6 ± 1.5, 17.4 ± 1.7 and 6.8 ± 0.7 % ID/g, respectively. The corresponding tumor uptake in mice coinjected with 0.05 mg cold bevacizumab were 5.8 ± 1.3, 8.9 ± 1.9 and 7.4 ± 1.0 % ID/g, respectively at the same time point, demonstrating specific blockage of the target in VEGF‐A–secreting tumors. The LS‐174T and SKOV3 tumors were clearly visualized by PET imaging after injecting 1.8–2.0 MBq 86Y‐CHX‐A″‐DTPA‐bevacizumab. Organ uptake quantified by PET closely correlated (r2 = 0.87, p = 0.64, n = 18) to values determined by biodistribution studies. This preclinical study demonstrates the potential of the radioimmunoconjugate, 86Y‐CHX‐A″‐DTPA‐bevacizumab, for noninvasive assessment of the VEGF‐A tumor angiogenesis status and as a surrogate marker for 90Y‐CHX‐A″‐DTPA‐bevacizumab radioimmunotherapy.
Curcumin enhances dasatinib‐induced inhibition of growth and transformation of colon cancer cells
Authors: Nautiyal Jyoti , Banerjee Sanjeev , Kanwar Shailender S. , Yu Yingjie , Patel Bhaumik B , Sarkar Fazlul H. , Majumdar Adhip P.N.
Published: April 2010

Colorectal cancer is the third most common form of malignancy, behind prostate and lung cancers. Despite recent advances in medicine, mortality from colorectal cancer remains high, highlighting the need for improved therapies. Numerous studies have demonstrated increased activation of EGFR and its family members (EGFRs), IGF‐1R as well as c‐Src in colorectal cancer. The current study was undertaken to examine the effectiveness of combination therapy of dasatinib (BMS‐354825; Bristol‐Myers Squibb), a highly specific inhibitor of Src family kinases (SFK) and a nontoxic dietary agent; curcumin (diferuloylmethane), in colorectal cancer in in vitro and in vivo experimental models. For the latter, we utilized C57BL/6 APCMin+/− mice. Initial in vitro studies revealed synergistic interactions between the two agents. Additionally, we have observed that combination treatment causes a much greater inhibition of the following metastatic processes than either agent alone: (i) colony formation, (ii) invasion through extracellular matrix and (iii) tubule formation by endothelial cells. Dasatinib affects the cell adhesion phenotype of colon cancer HCT‐116 cells whereas the combination therapy enhances this effect to a greater extent. Preclinical investigation revealed that the combination therapy to be highly effective causing an over 95% regression of intestinal adenomas in ApcMin+/− mice, which could be attributed to decreased proliferation and increased apoptosis. In conclusion, our data suggest that combination treatment of dasatinib and curcumin could be a potential therapeutic strategy for colorectal cancer.
Epstein‐Barr virus‐encoded EBNA‐5 forms trimolecular protein complexes with MDM2 and p53 and inhibits the transactivating function of p53
Authors: Kashuba Elena , Yurchenko Mariya , Yenamandra Surya Pavan , Snopok Boris , Szekely Laszlo , Bercovich Beatrice , Ciechanover Aaron , Klein George
Published: April 2010

We report that MDM2, a negative regulator of p53, can bind to EBNA‐5. Using GST pull‐down assay, immunoprecipitation, surface plasmon resonance and immunostaining of lymphoblastoid cells, we found that trimolecular complexes are formed between EBNA‐5, MDM2 and p53, where MDM2 serves as a bridge. The EBNA‐5 binding to MDM2 counteracted destabilizing effect of the latter on the p53. In ubiquitination and degradation assays in vitro, EBNA‐5 inhibited p53 polyubiquitination (but not monoubiquitination) in a concentration‐dependent manner. This resembles the effect of p14ARF on p53. Moreover, EBNA‐5 was found to inhibit the degradation of p53 in vitro. High levels of p53 expression were maintained in LCLs. The binding of EBNA‐5 to MDM2 also could impair the functional activity of p53. The p53‐dependent genes P21 and VDR were not induced in EBV‐infected, in contrast to mitogen‐activated cells. This may explain the tolerance of established LCLs to high levels of p53 without undergoing apoptosis.
Assessment of isorhamnetin 3‐O‐neohesperidoside from Acacia salicina: protective effects toward oxidation damage and genotoxicity induced by aflatoxin B1 and nifuroxazide
Authors: Bouhlel Ines , Limem Ilef , Skandrani Ines , Nefatti Aicha , Ghedira Kamel , Dijoux‐Franca Marie‐Genevieve , Leila Chekir‐Ghedira
Published: April 2010

Antioxidant activity of isorhamnetin 3‐O‐neohesperidoside, isolated from the leaves of Acacia salicina, was determined by the ability of this compound to inhibit xanthine oxidase activity and to scavenge the free radical 2,2′‐azino‐bis (3‐ethylbenzthiazoline‐6‐sulfonic acid) (ABTS.−) diammonium salt. Antigenotoxic activity was assessed using the SOS chromotest assay. This compound has the ability to scavenge the ABTS.+ radical by a hydrogen donating mechanism. We also envisaged the study of the antioxidant effect of this compound by the enzymatic xanthine/xanthine oxidase (X/XOD) assay. Results indicated that isorhamnetin 3‐O‐neohesperidoside was a potent inhibitor of xanthine oxidase and superoxide anion scavengers. Moreover, this compound induced an inhibitory activity against nifuroxazide and aflatoxine B1 (AFB1) induced genotoxicity. Taken together, these observations provide evidence that isorhamnetin 3‐O‐neohesperidoside isolated from the leaves of A. salicina is able to protect cells against the consequences of oxidative stress. Copyright © 2010 John Wiley & Sons, Ltd.
Bilateral round ligament varicosities mimicking an inguinal hernia in pregnancy: Case report
Authors: Kahriman Guven , Donmez Halil , Mavili Ertugrul , Ozcan Nevzat
Published: April 2010

A 22‐year‐old pregnant woman presented with a painful swelling in the right groin. Sonography was performed to confirm a presumptive diagnosis of inguinal hernia based on physical examination. Gray‐scale sonography examination revealed bilateral inguinal cystic lesion expanding with the Valsalva maneuver. Color Doppler imaging demonstrated multiple prominent vessels with retrograde venous flow during Valsalva maneuver. Bilateral round ligament varicosities were diagnosed and inguinal hernia was excluded by sonographic findings. Round ligament varicosities should be considered in the differential diagnosis of groin swelling during pregnancy. © 2010 Wiley Periodicals, Inc. J Clin Ultrasound 38:512–514, 2010
Sonographic appearance of a metastasis to the breast from a cerebellar medulloblastoma
Authors: Ternier Frédéric , Hadjaj Djamal , Jacquemier Jocelyne
Published: April 2010

We present the case of a 29‐year‐old woman with a medulloblastoma of the cerebellum who developed a breast mass during the course of her disease. Core biopsy of the breast lesion revealed a metastatic medulloblastoma. Development of metastasis to the breast from medulloblastoma is very rare and the prognosis for such patients is poor. It is important to distinguish primary breast cancer from metastasis to the breast, because the therapeutic options as well as the prognosis are very different. © 2010 Wiley Periodicals, Inc. J Clin Ultrasound, 2010
Sonographic diagnosis and successful medical management of an intramural ectopic pregnancy
Authors: Ong Clara , Su Lin‐Lin , Chia Dawn , Choolani Mahesh , Biswas Arijit
Published: April 2010

We report a case of posterior wall intramural pregnancy in a 36‐year‐old primigravida at 6 weeks gestation. Sonographic examination showed a viable intramural pregnancy distorting the contour of the uterus. The pregnancy was terminated by intrafetal injection of potassium chloride followed by intra‐amniotic methotrexate injection. © 2010 Wiley Periodicals, Inc. J Clin Ultrasound, 2010
Combined use of RNAi strategies with in vitro mouse limb bud culture
Authors: Zhu Yongfei , Zhu Jiangbo , Wan Xuying , Zhu Yuping , Zhang Tianbao
Published: April 2010

Methods for knocking out genes and generating transgenic animals are available to study gene functions during embryogenesis. Recently, RNA interference (RNAi), a conserved eukaryotic mechanism in which double‐stranded RNA induces sequence‐specific degradation of homologous mRNAs, has become a powerful tool for investigating gene function in cells, whole animals, and embryos. Additionally, in vitro mouse limb bud culture is available to study limb development. By microinjection, we established a new method combining the use of RNAi strategies with in vitro mouse limb bud culture to investigate the functions of various genes during embryonic limb development. Using this new method, we found that HspB10 may play a role during skeletal development of limbs. It shows that the combined use of RNAi strategies (via microinjection) with in vitro mouse limb bud culture system may become a technique for investigating gene function in limb development. J. Exp. Zool. (Mol. Dev. Evol.) 314B:412–416, 2010. © 2010 Wiley‐Liss, Inc.
Possibility of mixed progenitor cells in sea star arm regeneration
Authors: Hernroth Bodil , Farahani Farhad , Brunborg Gunnar , Dupont Sam , Dejmek Annika , Nilsson Sköld Helen
Published: April 2010

In contrast to most vertebrates, invertebrate deuterostome echinoderms, such as the sea star Asterias rubens, undergo regeneration of lost body parts. The current hypothesis suggests that differentiated cells are the main source for regenerating arm in sea stars, but there is little information regarding the origin and identity of these cells. Here, we show that several organs distant to the regenerating arm responded by proliferation, most significantly in the coelomic epithelium and larger cells of the pyloric caeca. Analyzing markers for proliferating cells and parameters indicating cell ageing, such as levels of DNA damage, pigment, and lipofuscin contents as well as telomere length and telomerase activity, we suggest that cells contributing to the new arm likely originate from progenitors rather than differentiated cells. This is the first study showing that cells of mixed origin may be recruited from more distant sources of stem/progenitor cells in a sea star, and the first described indication of a role for pyloric caeca in arm regeneration. Data on growth rate during arm regeneration further indicate that regeneration is at the expense of whole animal growth. We propose a new working hypothesis for arm regeneration in sea stars involving four phases: wound healing by coelomocytes, migration of distant progenitor cells of mixed origin including from pyloric caeca, proliferation in these organs to compensate for cell loss, and finally, local proliferation in the regenerating arm. J. Exp. Zool. (Mol. Dev. Evol.) 314B:457–468, 2010. © 2010 Wiley‐Liss, Inc.
Derivation of fluorine‐containing pyridinedicarboxylates. III Regio‐selective anion chemistry at the 2‐ and 4‐position
Authors: Chupp John P. , Molyneaux John M.
Published: April 2010

In contrast of Part II of the series, 4‐alkyl‐2‐(difluoromethyl)‐6‐(trifluoromethyl)‐3,5‐pyridinedicarboxylates, 1, were reacted with various strength bases to effect regio‐selective reaction of electrophiles at either the benzylic carbanion of the 4‐position, or at the 2‐(difluoromethyl)carbanion. Weaker bases up to and including potassium butoxide or lithium bis(trimethylsilylamide) effected reaction of 1 at the 4‐position to produce 2 and 3 by Stobbe‐type condensations of aldehydes and ketones. In similar manner carbon disulfide, carbon dioxide, alkyl halides, silyl halides, and hexachloroethane produced the highly functionalized derivatives 4‐10. In contrast, use of lithium diisopropylamide and like bases selectively effected carbanion formation at the 2‐position to form, with the cited electrophiles and others, substitution products 11. The latter were further derived to the highly functional materials, 12‐19.
Introduction to a culture, process, and philosophy
Authors: Bunting Jr. Robert F.
Published: April 2010

Patient‐ and family‐centered care is not a new concept, but it has garnered increased interest and support during the last decade. While most healthcare providers are excellent at providing clinical care, the practice of addressing the myriad other patient and family needs requires a conscious and collaborative effort. Though the staff of hospitals and other patient settings across the country have achieved various degrees of successful implementation of patient‐ and family‐centered care initiatives, the success has not been universal. An objective evaluation of patient‐ and family‐centered initiatives is needed. However, underdeveloped evaluation processes should not prevent the implementation of best practices that already have shown demonstrable success.
Enterprise‐wide effort brings patient perspective into mix
Authors: Sodomka Patricia , Spake Michael A. , Rush Jr. James J.
Published: April 2010

The National Patient Safety Foundation presented its 2009 Socius Award to the Medical College of Georgia (MCG Health). The award was given in recognition of the positive manner in which MCG Health collaborates with patients and families. The success achieved by MCG Health detailed in this article can motivate other hospital leaders, and the examples of collaboration can serve as templates for developing similar programs.
Building confidence into communication of bad news: The role of the patient advocate
Authors: Sullivan Ruth P. , Waldemayer Carol R. , Bunting Jr. Robert F.
Published: April 2010

The need for a patient advocate is greater than ever as medical errors continue to occur. News media quickly capture the egregious errors, but more errors are experienced by patients who suffer quietly. These patients know something wrong occurred during their hospitalization, but they choose to refrain from pursuing litigation against the providers. There also are thousands of individuals who never realize that a medical error occurred. In a patient‐ and family‐centered care environment, patient advocates can bridge these issues by participating on the healthcare team that is involved with the initial disclosure of the event and by providing a caring relationship to assure the patient's voice is heard and understood.
Epstein–Barr virus can infect rabbits by the intranasal or peroral route: An animal model for natural primary EBV infection in humans
Authors: Okuno Keisuke , Takashima Kazuaki , Kanai Kyosuke , Ohashi Makoto , Hyuga Ryosuke , Sugihara Hirotsugu , Kuwamoto Satoshi , Kato Masako , Sano Hitoshi , Sairenji Takeshi , Kanzaki Susumu , Hayashi Kazuhiko
Published: April 2010

Epstein–Barr virus (EBV) is spread universally in humans, and it causes infectious mononucleosis and sometimes induces serious EBV‐associated disease. The detailed mechanism of primary infection in humans has remained unclear, because it is difficult to examine the dynamics of EBV in vivo. In this study, a natural EBV‐infection rabbit model by intranasal or peroral inoculation is described. Ten male rabbits were examined for EBV‐DNA or mRNA expression and anti‐EBV antibodies in blood. Four of 10 rabbits showed the evidence of EBV infection; detection of EBV‐DNA or EBV‐related genes mRNA in peripheral blood mononuclear cells, increased EBV antibodies in the plasma, and the presence of lymphocytes expressing EBER1 and EBV‐related gene proteins in the lymphoid tissues of a rabbit. Three of four infected rabbits were detected transiently EBV‐DNA and/or mRNA of EBV‐related genes such as EBNA1, EBNA2, BZLF1, and EA in blood, while in one of four, EBV‐DNA and/or mRNA were detected for more than 200 days after viral inoculation. The level of EA‐IgG increased and its level was maintained in all infected rabbits, whereas those of VCA‐IgM and VCA‐IgG increased transiently, and EBNA‐IgG was not elevated. Pathological examination of a rabbit infected transiently revealed some scattered lymphocytes expressing EBER1, LMP1, and EBNA2 in the spleen and lymph nodes. EA expression was also observed in the spleen. These findings suggest that EBV can infect the rabbit by the intranasal or peroral route, and that this rabbit model is useful for examining the pathophysiology of natural primary EBV infection in humans. J. Med. Virol. 82:977–986, 2010. © 2010 Wiley‐Liss, Inc.
Do steatosis and steatohepatitis impact on sustained virological response (SVR) rates in patients receiving pegylated interferon and ribavirin for chronic hepatitis C infection?
Authors: Cross T.J.S. , Quaglia A. , Nolan J. , Hughes S. , Harrison P.M.
Published: April 2010

The impact of steatosis on treatment response in chronic hepatitis C infection is controversial. The aim of this study was to determine whether steatosis ± steatohepatitis on pre‐treatment liver biopsy influenced sustained virological response (HCV RNA negative 6 months after completing therapy) in patients with chronic hepatitis C infection treated with pegylated interferon‐α and ribavirin. One hundred and seventy‐nine patients, median age 46 years (interquartile range 40–52), treated between 2001 and 2005. Histological evidence of steatosis was present in 93 patients (52%) and steatohepatitis in 33 patients (18%), 31 patients (17.3%) were cirrhotic. There were 106 (59%) responders, who were similar to non‐responders in respect to gender, age, and pre‐treatment ALT. On univariate analysis, infection with genotype 2 or 3 was associated with sustained virological response (odds ratio 6.5 (95% CI, 3.3–12.5); P < 0.0001), whereas cirrhosis and patient weight were associated with a reduced response (odds ratios 0.23 (95% CI, 0.11–0.48); P < 0.0001, and 0.97 (95% CI, 0.95–0.99); P < 0.01, respectively); steatohepatitis but not steatosis impacted on the likelihood of achieving sustained virological response (odds ratio 0.37 (95% CI, 0.17–0.77); P = 0.009, and P = 0.18, respectively). Multivariate analysis revealed that infection with genotype 1 or 4 (odds ratio 0.09 (95% CI, 0.03–0.32); P < 0.001) and pre‐treatment weight (odds ratio 0.94 (95% CI, 0.90–0.98); P = 0.002) were the only variables associated independently with sustained virological response. In chronic hepatitis C infection, although steatosis was associated with steatohepatitis, neither was shown to affect sustained virological response, which was influenced by genotype, patient weight and the presence of cirrhosis. J. Med. Virol. 82:958–964, 2010. © 2010 Wiley‐Liss, Inc.