bluuebunny/crossref_metadata_2025 huggingface dataset:
Therapeutic effect of recombinant human epidermal growth factor (rhEGF) on mucositis in patients undergoing radiotherapy, with or without chemotherapy, for head and neck cancer
Authors: Wu Hong Gyun , Song Si Yeol , Kim Yeon Sil , Oh Young Taek , Lee Chang Geol , Keum Ki Chang , Ahn Yong Chan , Lee Sang‐wook
Published: June 2009

BACKGROUND:

We evaluated the efficacy of topically applied human recombinant epidermal growth factor (rhEGF) for the treatment of oral mucositis induced by radiotherapy (RT), with or without chemotherapy, in patients with head and neck cancer.

METHODS:

Patients receiving definitive chemoradiotherapy, definitive RT, or postoperative RT to the oral cavity or oropharynx were recruited from 6 institutions and enrolled in a randomized, double‐blind, placebo‐controlled phase 2 trial. Patients were assigned to a placebo group or to 1 of 3 EGF‐treatment groups (10, 50, or 100 μg/mL doses, delivered in a spray, twice daily). The grade of mucositis was evaluated using the Radiation Therapy Oncology Group (RTOG) scoring criteria. Responders to EGF were defined as having an RTOG grade of 2 or lower at the fourth‐ or fifth‐week examinations during RT, but an enduring RTOG grade 2 for 2 weeks was an exception.

RESULTS:

Of the 113 patients included in the study, 28 received placebo and 29 received EGF at 10 μg/mL, 29 at 50 μg/mL, and 27 at 100 μg/mL. EGF significantly reduced the incidence of severe oral mucositis at the primary endpoint (a 64% response was observed with 50 μg/mL EGF vs a 37% response in the control group; P = .0246).

CONCLUSIONS:

The EGF oral spray may have potential benefit for oral mucositis in patients undergoing RT for head and neck cancer. Phase 3 studies are ongoing to confirm these results. Cancer 2009. © 2009 American Cancer Society.
Candidate therapeutic agents for hepatocellular cancer can be identified from phenotype‐associated gene expression signatures
Authors: Braconi Chiara , Meng Fanyin , Swenson Erica , Khrapenko Lyudmyla , Huang Nianyuan , Patel Tushar
Published: June 2009

BACKGROUND:

The presence of vascular invasion in hepatocellular cancer (HCC) correlates with prognosis, and is a critical determinant of both the therapeutic approach and the recurrence or intrahepatic metastases. The authors sought to identify candidate therapeutic agents capable of targeting the invasive phenotype in HCC.

METHODS:

A gene expression signature associated with vascular invasion derived from 81 human cases of HCC was used to screen a database of 453 genomic profiles associated with 164 bioactive molecules using the connectivity map. Candidate agents were identified by their inverse correlation to the query gene signature. The efficacy of the candidate agents to target invasion was experimentally verified in PLC/PRF‐5 and HepG2 HCC cells.

RESULTS:

The gene signature associated with vascular invasion in HCC comprised of 47 up‐regulated and 26 down‐regulated genes. Computational bioinformatics analysis revealed several putative candidates, including resveratrol and 17‐allylamino‐geldanamycin (17‐AAG). Both of these agents reduced HCC cell invasion at noncytotoxic concentrations. 17‐AAG, a heat shock protein 90 (HSP‐90) inhibitor, was shown to modulate the expression of several diverse cancer‐associated genes, including ADAMTS1, part of the query signature, and maspin, an HSP‐90–associated protein with a tumor suppressor role in HCC.

CONCLUSIONS:

Candidates for further evaluation as therapies to limit invasion in HCC have been identified using a computational bioinformatics analysis of phenotype‐associated gene expression. Phenotype targeting using genomic profiling is a rational approach for drug discovery. Therapeutic strategies targeting a defined cancer‐associated phenotype can be identified without a detailed knowledge of individual downstream targets. Cancer 2009. © 2009 American Cancer Society.
Significance of suboptimal response to imatinib, as defined by the European LeukemiaNet, in the long‐term outcome of patients with early chronic myeloid leukemia in chronic phase
Authors: Alvarado Yesid , Kantarjian Hagop , O'Brien Susan , Faderl Stefan , Borthakur Gautam , Burger Jan , Wierda William , Garcia‐Manero Guillermo , Shan Jianqin , Cortes Jorge
Published: June 2009

BACKGROUND:

The European LeukemiaNet recommendations for chronic myeloid leukemia (CML) defined a group of patients with suboptimal response to imatinib. The significance of this response was not well defined.

METHODS:

The significance of having had a suboptimal response during imatinib therapy among 281 patients with CML treated with standard‐dose (n = 73) or high‐dose (n = 208) imatinib was investigated.

RESULTS:

Rates of suboptimal response at 6, 12, and 18 months were 4%, 8%, and 40%, respectively, and were not influenced by Sokal risk score. Patients with a suboptimal response at 6 months had a significantly lower probability of eventually achieving a complete cytogenetic response (CCyR) compared with those with an optimal response (30% vs 97%; P < .001), and their event‐free survival (EFS) and transformation‐free survival (TFS) were found to be similar to those with criteria for failure at this time point. Suboptimal response at 12 months defined a group with a similar TFS as those with optimal response, but with worse EFS. In contrast, patients with a suboptimal response at 18 months had outcomes that were similar to those patients with an optimal response. A multivariate analysis confirmed the significance of response category after adjusting for pretreatment characteristics and imatinib dose.

CONCLUSIONS:

The results of the current study suggested that suboptimal response was a heterogeneous category, and some patients had an outcome that mirrored that of patients with failed therapy. Interventions aimed at improving this outcome are required. Cancer 2009. © 2009 American Cancer Society.
Dynamics and management of cytopenias associated with dasatinib therapy in patients with chronic myeloid leukemia in chronic phase after imatinib failure
Authors: Quintás‐Cardama Alfonso , De Souza Santos Fabio Pires , Kantarjian Hagop , O'Brien Susan , Faderl Stefan , Awais Ahmed , Borthakur Gautam , Cortes Jorge
Published: June 2009

BACKGROUND:

The incidence, dynamics, and management of cytopenias were investigated in patients with chronic myeloid leukemia in chronic phase (CP CML) who received dasatinib therapy after imatinib failure.

METHODS:

Data were analyzed from 130 patients with CP CML who were treated with dasatinib from November 2003 to March 2006 in phase 1 (n = 22) or phase 2 or 3 (n = 108) studies for the development of grade 2 to 4 cytopenia (according to the National Cancer Institute Common Terminology Criteria [version 3.0]).

RESULTS:

Grade 2 to 4 neutropenia and/or thrombocytopenia occurred in 94 (72%) patients during dasatinib therapy and grade 3 to 4 occurred in 67 (52%) patients. Of the 94 patients who developed grade 2 to 4 neutropenia and/or thrombocytopenia, 64 (68%) also developed at least grade 2 anemia, and 16 (17%) developed grade 3 to 4 anemia. Management of cytopenias included transient dasatinib interruption in 35 (37%) patients, filgrastim in 12 (14%) patients, recombinant erythropoietin in 29 (45%) patients, and interleukin‐11 in 3 (5%) patients. Factors associated with an increased risk for developing grade 2 to 4 cytopenias were longer time from diagnosis to treatment, prior interferon or imatinib therapy, and a lower white blood cell count at the initiation of dasatinib therapy.

CONCLUSIONS:

Hematologic toxicity was frequent during dasatinib therapy in patients with CP CML, particularly at doses >100 mg daily. Treatment interruption and/or dose reduction as well as growth factor support were found to be safe and efficacious strategies to facilitate the continuous administration of dasatinib. Cancer 2009. © 2009 American Cancer Society.
Pancreatic resection
Authors: Murphy Melissa M. , Simons Jessica P. , Hill Joshua S. , McDade Theodore P. , Chau Ng Sing , Whalen Giles F. , Shah Shimul A. , Harrison Lynn H. , Tseng Jennifer F.
Published: June 2009

BACKGROUND:

Blacks are affected disproportionately by pancreatic adenocarcinoma and have been linked with poor survival. Surgical resection remains the only potential curative option. If surgical disparities exist, then they may provide insight into outcome discrepancies.

METHODS:

Patients with pancreatic adenocarcinoma were identified using the National Cancer Institute's Surveillance, Epidemiology, and End Results data from 1992 to 2002. Univariate analyses were used to compare demographics, tumor characteristics, and surgical data; and logistic regression was used to determine independent predictors for recommendation/performance of surgery. Kaplan‐Meier survival was assessed, and a Cox proportional hazards model was used to examine adjusted predictors of survival.

RESULTS:

In total, 27,828 patients were identified; 81.4% were white, 11.5% were black, 7.2% were of other race. White patients and black patients presented with similar stage and had surgery recommended at similar rates (34.5% vs 34%, respectively; P = .57). Black patients underwent fewer resections (10.6% vs 12.7%; P < .001). Multivariate analysis confirmed that black patients were less likely to undergo resection (adjusted odds ratio, 0.69; 95% confidence interval [95% CI], 0.57‐0.84). Overall, black patients had worse univariate survival. The survival among black patients who underwent resection did not differ statistically from the survival of similar white patients, although the median survival trended lower (11 months vs 13 months; P = .13). In a multivariate Cox model, black race predicted worse survival (hazards ratio, 1.11; 95% CI, 1.07‐1.16), and pancreatic resection was protective (hazards ratio, 0.56; 95% CI, 0.53‐0.59).

CONCLUSIONS:

Black and white patients with pancreatic adenocarcinoma presented with similar stages and were recommended for pancreatectomy at similar rates, yet black patients underwent fewer resections. After resection, crude survival did not differ significantly between white and black patients, although multivariate analysis demonstrated a survival disadvantage for blacks despite adjusting for resection. The current results suggested that pancreatectomy may be underused for blacks. Maximizing resection rates for appropriate patients may be an important component in reducing outcome disparities for pancreatic adenocarcinoma. Cancer 2009. © 2009 American Cancer Society.
Correlative analysis of gene expression profile and prognosis in patients with gliomatosis cerebri
Authors: D'Urso Oscar Fernando , D'Urso Pietro Ivo , Marsigliante Santo , Storelli Carlo , Luzi Giuseppe , Gianfreda Cosimo Damiano , Montinaro Antonio , Distante Alessandro , Ciappetta Pasqualino
Published: June 2009

BACKGROUND:

In modern clinical neuro‐oncology, the pathologic diagnoses are very challenging, creating significant clinical confusion and affecting therapeutic decisions and prognosis.

METHODS:

TP53 and PTEN gene sequences were analyzed, and microarray expression profiling was also performed. The authors investigated whether gene expression profiling, coupled with class prediction methodology, could be used to determine the prognosis of gliomatosis cerebri in a more consistent manner than standard pathology.

RESULTS:

The authors reported the results of a molecular study in 59 cases of gliomatosis cerebri, correlating these results with prognosis. The well‐known prognostic factors of gliomas (ie, age, Karnofsky performance status, histology [grade 2 vs 3], and contrast enhancement) were found to be predictive of response or outcome in only a percentage of patients but not in all patients. The authors identified a 23‐gene signature that was able to predict patient prognosis with microarray gene expression profiling. With the aim of producing a prognosis tool that is useful in clinical investigation, the authors studied the expression of this 23‐gene signature by real‐time quantitative polymerase chain reaction. Real‐time expression values relative to these 23 gene features were used to build a prediction method able to distinguish patients with a good prognosis (those more likely to be responsive to therapy) from patients with a poor prognosis (those less likely to be responsive to therapy).

CONCLUSIONS:

The results of the current study demonstrated not only a strong association between gene expression patterns and patient survival, but also a robust replicability of these gene expression–based predictors. Cancer 2009. © 2009 American Cancer Society.
Developing quality indicators for cancer end‐of‐life care
Authors: Seow Hsien , Snyder Claire F. , Shugarman Lisa R. , Mularski Richard A. , Kutner Jean S. , Lorenz Karl A. , Wu Albert W. , Dy Sydney M.
Published: June 2009

Quality indicators applicable to cancer end‐of‐life care exist, but have not been widely implemented. To advance this field, the authors worked with the Agency for Health Care Research and Quality and the National Cancer Institute to organize a national symposium to discuss key issues and future goals, based on a conceptual framework. Discussions focused on 8 key domains in end‐of‐life cancer care: pain; dyspnea; communication, care planning, and decision making; psychosocial care; communication about chemotherapy; depression; continuity, coordination, and care transitions; and spirituality and closure. Key themes included the need for clarity on definitions and key aspects of care within domains, the need to start implementing indicators in more developed domains, and the importance of high‐quality symptom assessment and documentation of key processes. Key areas for future work include development of more outcome indicators, methods to better incorporate indicators and patient‐reported outcomes into clinical processes of care, and coordination across domains and settings. Measuring the quality of end‐of‐life cancer care is essential to understanding how best to improve patient outcomes and care. Cancer 2009. © 2009 American Cancer Society.
Assessment of the performance of the Stanford Online Calculator for the prediction of nonsentinel lymph node metastasis in sentinel lymph node‐positive breast cancer patients
Authors: Scow Jeffrey S. , Degnim Amy C. , Hoskin Tanya L. , Reynolds Carol , Boughey Judy C.
Published: June 2009

BACKGROUND:

Several models for the prediction of nonsentinel lymph node (NSLN) metastasis in sentinel lymph node (SLN)‐positive breast cancer patients have been proposed. In this study, the authors evaluate the Stanford Online Calculator (SOC), which was designed to predict the likelihood of NSLN metastasis using only 3 variables: primary tumor size, SLN metastasis size, and angiolymphatic invasion status. They compared it with the Mayo and Memorial Sloan‐Kettering Cancer Center (MSKCC) nomograms.

METHODS:

The SOC was used to calculate the probability of NSLN metastasis in 464 breast cancer patients with SLN metastasis who underwent completion axillary lymph node dissection at the Mayo Clinic. The area under the receiver operating characteristic curve (AUC) was calculated for each model. Mean probabilities of patients with and without NSLN metastasis were compared. Patients with ≤5%, ≤10%, and 100% NSLN metastasis probabilities were examined.

RESULTS:

The AUCs of the Stanford, MSKCC, and Mayo models were 0.72, 0.74, and 0.77, respectively (P = .13). The mean Stanford probabilities for patients with and without NSLN metastasis were 0.75 (range, 0.06‐1.0) and 0.50 (range, 0.05‐1.0), respectively (P < .0001). The false‐negative rates for patients with a Stanford probability of ≤5% and ≤10% were 0% and 13%, respectively. Of the patients with a Stanford probability of 100%, 26% did not have NSLN metastasis.

CONCLUSIONS:

Despite using only 3 variables, the Stanford nomogram appears to perform on a par with, but not better than, the MSKCC and Mayo nomograms. Further validation in other patient populations is needed. Cancer 2009. © 2009 American Cancer Society.
Transgenesis in Xenopus using the Sleeping Beauty transposon system
Authors: Yergeau Donald A. , Johnson Hamlet Michelle R. , Kuliyev Emin , Zhu Haiqing , Doherty Joanne R. , Archer Taylor D. , Subhawong Andrea P. , Valentine Marc B. , Kelley Clair M. , Mead Paul E.
Published: June 2009

Transposon‐based integration systems have been widely used for genetic manipulation of invertebrate and plant model systems. In the past decade, these powerful tools have begun to be used in vertebrates for transgenesis, insertional mutagenesis, and gene therapy applications. Sleeping Beauty (SB) is a member of Tc1/mariner class of transposases and is derived from an inactive form of the gene isolated from Atlantic salmon. SB has been used extensively in human cell lines and in whole animal vertebrate model systems such as the mouse, rat, and zebrafish. In this study, we describe the use of SB in the diploid frog Xenopus tropicalis to generate stable transgenic lines. SB transposon transgenes integrate into the X. tropicalis genome by a noncanonical process and are passed through the germline. We compare the activity of SB in this model organism with that of Tol2, a hAT (hobo, Ac1, TAM)‐like transposon system. Developmental Dynamics 238:1727–1743, 2009. © 2009 Wiley‐Liss, Inc.
Rnf19a, a ubiquitin protein ligase, and Psmc3, a component of the 26S proteasome, Tether to the acrosome membranes and the head–tail coupling apparatus during rat spermatid development
Authors: Rivkin Eugene , Kierszenbaum Abraham L. , Gil Mara , Tres Laura L.
Published: June 2009

We report the cDNA cloning of rat testis Rnf19a, a ubiquitin protein ligase, and show 98% and 93% protein sequence identity of testicular mouse and human Rnf19a, respectively. Rnf19a interacts with Psmc3, a protein component of the 19S regulatory cap of the 26S proteasome. During spermatid development, Rnf19a and Psmc3 are initially found in Golgi‐derived proacrosomal vesicles. Later on, Rnf19a, Psmc3, and ubiquitin are seen along the cytosolic side of the acrosomal membranes and the acroplaxome, a cytoskeletal plate linking the acrosome to the spermatid nuclear envelope. Rnf19a and Psmc3 accumulate at the acroplaxome marginal ring–manchette perinuclear ring region during spermatid head shaping and in the developing sperm head–tail coupling apparatus and tail. Rnf19a and Psmc3 may interact directly or indirectly with each other, presumably pointing to the participation of the ubiquitin–proteasome system in acrosome biogenesis, spermatid head shaping, and development of the head‐tail coupling apparatus and tail. Developmental Dynamics 238:1851–1861, 2009. © 2009 Wiley‐Liss, Inc.
Microinjection of Cre recombinase protein into zygotes enables specific deletion of two eukaryotic selection cassettes and enhances the expression of a DsRed2 reporter gene in Ccr2/Ccr5 double‐deficient mice
Authors: Luckow Bruno , Hänggli Amy , Maier Holger , Chilla Silvia , Loewe Robert P. , Dehmel Stefan , Schlöndorff Detlef , Loetscher Pius , Zerwes Hans‐Günter , Müller Matthias
Published: June 2009

The chemokine receptors CCR2 and CCR5 represent potential novel therapeutic targets to treat important inflammatory and infectious diseases, including atherosclerosis and HIV infection. To study the functions of both receptors in vivo, we aimed to generate Ccr2/Ccr5 double‐deficient mice. As these genes are separated by <20 kb, they were inactivated consecutively by two rounds of gene targeting in embryonic stem (ES) cells. Thereby neomycin and hygromycin selection cassettes flanked by four identical loxP recognition sequences for Cre recombinase were integrated into the ES cell genome together with EGFP and DsRed2 reporter genes. Both selection cassettes could be deleted in vitro by transiently transfecting ES cells with Cre expression vectors. However, after blastocyst microinjection these cells yielded only weak chimeras, and germline transmission was not achieved. Therefore, Ccr2/Ccr5 double‐deficient mice were generated from ES cells still carrying both selection cassettes. Microinjection of zygotes with a recombinant fusion protein consisting of maltose‐binding protein and Cre (MBP‐Cre) allowed the selective deletion of both cassettes. All sequences in between and both reporter genes were left intact. Deletion of both selection cassettes resulted in enhanced DsRed2 reporter gene expression. Cre protein microinjection of zygotes represents a novel approach to perform complex recombination tasks. genesis 47:545–558, 2009. © 2009 Wiley‐Liss, Inc.
Commentaries on ‘Abstinence‐plus programs for HIV infection prevention in high‐income countries’
Authors: Flicker Sarah , Guilamo‐Ramos Vincent , Bouris Alida
Published: June 2009

These are commentaries on a Cochrane review, published in this issue of EBCH, first published as: Underhill K, Montgomery P, Operario D. Abstinence‐plus programs for HIV infection prevention in high‐income countries. Cochrane Database of Systematic Reviews 2008, Issue 1. Art. No.: CD007006. DOI: 10.1002/14651858.CD007006.

Further information for this Cochrane review is available in this issue of EBCH in the accompanying Summary article. Copyright © 2009 The Cochrane Collaboration. Published by John Wiley & Sons, Ltd. The Cochrane Collaboration
Two commentaries on ‘Family‐based programmes for preventing smoking by children and adolescents’
Authors: Priest Naomi , Waters Elizabeth , Norman Cameron D. , Best Allan
Published: June 2009

These are commentaries on a Cochrane review, published in this issue of EBCH, first published as: Thomas RE, Baker PRA, Lorenzetti D. Family‐based programmes for preventing smoking by children and adolescents. Cochrane Database of Systematic Reviews 2007, Issue 1. Art. No.: CD004493. DOI: 10.1002/14 651 858.CD004493.pub2.

Further information for this Cochrane review is available in this issue of EBCH in the accompanying Summary article. Copyright © 2009 The Cochrane Collaboration. Published by John Wiley & Sons, Ltd. The Cochrane Collaboration
Ilicic Acid as a Natural Quiron for the Efficient Preparation of Bioactive α‐ and β‐Eudesmol
Authors: Barrero Alejandro F. , Herrador M. Mar , Arteaga Pilar , Catalán Julieta V.
Published: June 2009

An efficient procedure for the isolation of the sesquiterpene ilicic acid (3) on a multigram scale of extracts obtained from aerial parts of Inula viscosa (Asteraceae) was developed. Acid 3 is an appropriate starting material for short, enantiospecific syntheses of β‐eudesmol (1) and α‐eudesmol (2), natural products featuring significant antiangiogenic and anti‐Alzheimer properties. Synthesis of 1 was achieved in six steps and the synthesis of 2 in seven, producing overall yields of 52 and 41 %, respectively.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Reagentless Biosensor for Hydrogen Peroxide Based on the Immobilization of Hemoglobin in Platinum Nanoparticles Enhanced Poly(chloromethyl thiirane) Cross‐linked Chitosan Hybrid Film
Authors: Jia Shanshan , Fei Junjie , Tian Tian , Zhou Fangqin
Published: June 2009

An unmediated hydrogen peroxide (H2O2) biosensor was prepared by co‐immobilizing hemoglobin (Hb) with platinum nanoparticles enhanced poly(chloromethyl thiirane) cross‐linked chitosan (CCCS‐PNs) hybrid film. CCCS could provide a biocompatible microenvironment for Hb and PNs could accelerate the electron transfer between Hb and the electrode. Spectroscopic analysis indicated that the immobilized Hb could maintain its native structure in the CCCS‐PNs hybrid film. Entrapped Hb exhibited direct electrochemistry for its heme Fe(III)/Fe(II) redox couples at −0.396 V in the CCCS‐PNs hybrid film, as well as peroxidase‐like activity to the reduction of hydrogen peroxide without the aid of an electron mediator.
Fabrication of Redox‐Mediator Supported Potentiometric Nitrate Biosensor with Nitrate Reductase
Authors: Sohail Manzar , Adeloju Samuel B.
Published: June 2009

Fabrication of a more superior nitrate potentiometric biosensor than previously achieved with NaR and NADH has been accomplished by co‐entrapment of redox mediators and NaR into polypyrrole (PPy) film during galvanostatic polymerization of pyrrole. The replacement of NADH with redox mediators such as thionin acetate (ThAc), safranin (Saf), and azure A (AzA) gave more sensitive potentiometric responses, better minimum detectable concentration, linear concentration range and response time for nitrate than possible with NADH. The co‐entrapment of ThAc, Saf, AzA and methyl viologen (MV) with NaR into PPy films also improved the Nernstian behavior of the electrode process beyond the capability of the PPy‐NaR‐NADH biosensor. Substantial reduction in volume and quantity of cofactor/mediator and, hence cost, was achieved by the replacement of NADH with a redox mediator. Only 50 μM of AzA was required to form a PPy‐NaR‐AzA biosensor which gave the most sensitive potentiometric response for nitrate, achieving a minimum detectable concentration of 10 μM, a linear concentration range of 50–5000 μM and a response time of 2–4 s.
Comparison of thermal‐ and photo‐polymerization of lauryl methacrylate monolithic columns for CEC
Authors: Bernabé‐Zafón Virginia , Cantó‐Mirapeix Amparo , Simó‐Alfonso Ernesto F. , Ramis‐Ramos Guillermo , Herrero‐Martínez José Manuel
Published: June 2009

Lauryl methacrylate‐based (LMA) monolithic columns for CEC, prepared using either thermal initiation or by UV‐irradiation in the presence of AIBN have been compared. Thermal polymerization was carried out at 70°C for 20 h. For UV initiation, the effects of the time exposure to UV light and irradiation energy were investigated. For each initiation process, the influence of composition of porogenic solvent (1,4‐butanediol/1‐propanol ratio) on the physical and electrochromatographic properties of the resulting monoliths was also evaluated. Photochemically lauryl methacrylate stationary phases initiated showed higher permeabilities and better efficiencies than those prepared by thermal initiation. After optimization of polymerization mixture, photopolymerized columns provided a permeability of 4.25×10−13 m2 and a minimum plate height of 13.4 μm for a mixture of polycyclic aromatic hydrocarbons. Similar column‐to‐column and batch‐to‐batch reproducibilities, with RSD values below 11.6 and 11.0 % for the thermal‐ and UV‐initiated columns, respectively, were obtained.
Improved dynamic range of protein quantification in silver‐stained gels by modelling gel images over time
Authors: Grove Harald , Færgestad Ellen M. , Hollung Kristin , Martens Harald
Published: June 2009

Silver staining is a commonly used protein stain to visualise proteins separated by 2‐DE. Despite this, the technique suffers from a limited dynamic range, making the simultaneous quantification of high‐ and low‐abundant proteins difficult. In this paper we take advantage of the fact that silver staining is not an end‐point stain by photographing the gels during development. This procedure provides information about the change in measured absorbance for each pixel in the protein spots on the gel. The maximum rate of change was found to be correlated with the amount of applied protein, providing a new way of estimating protein amount in 2‐DE gels. We observed an improvement in the dynamic range of silver staining by up to two orders of magnitude.
Online preconcentration and two‐dimensional separation of cationic compounds via hyphenation of capillary zone electrophoresis with cyclodextrin‐modified micellar electrokinetic capillary chromatography
Authors: Zhang Zhao‐Xiang , Zhang Ming‐Zhe , Zhang Shu‐Sheng
Published: June 2009

A novel preconcentration/separation approach, which online combined CZE with CD‐modified MEKC, was developed for simultaneous enhancing resolving power and detection sensitivity. CZE with cation‐selective exhaustive injection and transient ITP preconcentration was used as the first dimension, from which the effluent fractions were further analyzed by CD‐modified MEKC acting as the second dimension. As the key to successful integration of CZE with MEKC, a new interface was designed and electroaccumulation focusing strategy was employed to avoid analyte band diffusion at the interface. The comprehensive 2‐D system was successfully established with only one high voltage and four electrodes. The grouping of two orthogonal separation techniques, together with analytes preconcentration techniques, significantly enhanced resolution and sensitivity for 2‐D separation of cationic compounds. The resulting electrophoregram was quite different from that of either single CZE or MEKC. Up to 14 000‐ to 35 000‐fold improvement in sensitivity was obtained relative to conventional electrokinetic injection method. The limits of detection (S/N=3) were in the range of 0.03–0.1 μg/L. The number of theoretical plates was in the range of 103 000–184 000. This method was successfully applied to the analysis of trace cationic cardiovascular drugs in wastewater.
A simple preparative free‐flow electrophoresis joined with gratis gravity: I. Gas cushion injector and self‐balance collector instead of multiple channel pump
Authors: Chen Su , Palmer James F. , Zhang Wei , Shao Jing , Li Si , Fan Liu‐Yin , Sun Ren , Dong Yu‐Chao , Cao Cheng‐Xi
Published: June 2009

This paper describes a novel free‐flow electrophoresis (FFE), which is joined with gratis gravity, gas cushion injector (GCI) and self‐balance collector instead of multiple channel pump, for the purpose of preparative purification. The FFE was evaluated by systemic experiments. The results manifest that (i) even though one‐channel peristaltic pump is used for the driving of background buffer, there is still stable flow in the FFE chamber; (ii) the stable flow is induced by the gravity‐induced pressure due to the difference of buffer surfaces in the GCI and self‐balance collector; (iii) the pulse flow of background buffer induced by the peristaltic pump is greatly reduced by the GCI with good compressibility of included air; (iv) the FFE can be well used for zone electrophoretic separation of amino acids; (v) up to 20 inlets simultaneous sample injection and up to five to tenfold condensation of amino acid can be achieved by combining the FFE device with the method of moving reaction boundary. To the best of authors' knowledge, FFE has not been used for such separation and condensation of amino acids. The relevant results achieved in the paper have evident significance for the development of preparative FFE.
A new route for simple and rapid determination of hydrogen peroxide in RAW264.7 macrophages by microchip electrophoresis
Authors: Gong Xiaocong , Li Qingling , Xu Kehua , Liu Xin , Li Hongmin , Chen Zhenzhen , Tong Lili , Tang Bo , Zhong Hongbo
Published: June 2009

A new method using MCE with LIF detection was developed for the determination of hydrogen peroxide (H2O2). Bis(p‐methylbenzenesulfonyl)dichlorofluorescein, a new fluorogenic reagent synthesized by our laboratory was employed as a labeling reagent, the derivatization reaction was performed in 0.10 M HEPES buffer (pH 7.4) for 30 min at 37°C. The detection of H2O2 was accomplished in 55 s, using a 40 mM HEPES buffer, 20% mannitol, pH 7.4, on a glass microchip. The RSDs of migration time and peak area were 1.8 and 3.7%, respectively. Method validation showed the linear response ranging from 0.50 to 50 μM with an LOD (S/N=3) of 0.20 μM (19.1 amol). The proposed method was applied to determine H2O2 in phorbol myristate acetate‐stimulated RAW264.7 macrophages, the concentration of H2O2 was found to be 1.86±0.05 μM; recoveries for macrophage samples were from 96.7 to 97.8%, within‐days and between‐days accuracies were 4.5% (n=5) and 6.8% (n=5), respectively.
Characterisation of mixed lithium dodecyl sulphate/lithium perfluorooctanesulphonate pseudo‐stationary phases in MEKC
Authors: Varga András , Huszár Mónika , Dobos Zsófia , Kiss Éva , Horváth Anikó , Idei Miklós
Published: June 2009

Lipophilicity and methylene selectivity of mixed pseudo‐stationary phases (PSPs) (containing lithium dodecyl sulphate (LDS) and lithium perfluorooctanesulphonate (LiPFOS) in different molar ratios) applied in MEKC have been investigated. Micellar proportion (tprop,mic, a quantity expressing that how much time is spent by the analyte in the micellar phase related to its whole migration time), CLOGP50 value (showing the value of hydrophobicity of a molecule spending exactly 50% of its migration time in the PSP) and methylene selectivity have been determined for different LDS/LiPFOS mixed phases. Values of the above‐mentioned parameters have been determined for analytes with different chemical structures (alkylbenzene and alkylphenone homologous series, alcohols). Good linear correlation was obtained between either the micellar proportion, CLOGP50, or methylene selectivity and the phase composition for the mixed phases. Lipophilicity and methylene selectivity of the mixed LDS/LiPFOS PSPs can be calculated and can continuously be changed by mixing the two single phases (LDS and LiPFOS) in the appropriate (and calculable) portion.
Influence of polyelectrolyte capillary coating conditions on protein analysis in CE
Authors: Nehmé Reine , Perrin Catherine , Cottet Hervé , Blanchin Marie‐Dominique , Fabre Huguette
Published: June 2009

CE of biomolecules is limited by analyte adsorption on the capillary wall. To prevent this, monolayer or successive multiple ionic‐polymer layers (SMILs) of highly charged polyelectrolytes can be physically adsorbed on the inner capillary surface. Although these coatings have become commonly used in CE, no systematic investigation of their performance under different coating conditions has been carried out so far. In a previous study (Nehmé, R., Perrin, C., Cottet, H., Blanchin, M. D., Fabre, H., Electrophoresis 2008, 29, 3013–3023), we investigated the influence of different experimental parameters on coating stability, repeatability and peptide peak efficiency. Optimal coating conditions for monolayer and multilayer (SMILs) poly(diallyldimethylammonium) chloride/ poly(sodium 4‐styrenesulfonate) coated capillaries were determined. In this study, the influence of polyelectrolyte concentration and ionic strength of the coating solutions, and the number of coating layers on coating stability and performance in limiting protein adsorption was carried out. EOF magnitude and repeatability were used to monitor coating stability. Coating ability to limit protein adsorption was investigated by monitoring variations of migration times, time‐corrected peak areas and separation efficiency of test proteins. The separation performance of polyelectrolyte coatings were compared with those obtained with bare silica capillaries.
In‐capillary microextraction using monolithic polymers: Application to preconcentration of carbamate pesticides prior to their separation by MEKC
Authors: Rodríguez‐Gonzalo Encarnación , Ruano‐Miguel Luis , Carabias‐Martínez Rita
Published: June 2009

In the present work we report a novel procedure for in‐capillary microextraction using a monolithic polymeric sorbent. In the proposed methodology, sample treatment takes place in the CE instrument but in a different capillary from that used for the electrophoretic separation. Polymers based on butyl methacrylate and divinylbenzene formed in situ inside a capillary column were assayed. The best results were found with the divinylbenzene‐based polymers. The usefulness of the proposed procedure was checked for the determination of carbamate pesticides (Methomyl, Asulam, Carbendazim, Aldicarb, Carbetamide, Propoxur, Pirimicarb, Carbaryl, Carbofuran and Methiocarb) and three of their degradation compounds (Aldicarb sulphoxide, 2‐isopropoxyphenol and α‐naphthol) using MEKC. The optimization of the MEKC is reported, a good separation of the 13 analytes being obtained in less than 6 min. The analytical method using in‐capillary microextraction was validated in terms of linearity, repeatability, precision (RSD≤18% for 50 μg/L), and LODs (1–16 μg/L), and it revealed the usefulness of this in‐capillary preconcentration procedure for the determination of analytes of intermediate polarity.
Assessment of the finite volume method applied to the v2 − f model
Authors: Mortensen Mikael , Reif Bjørn Anders Pettersson , Wasberg Carl Erik
Published: June 2009

The objective of this paper is to assess the accuracy of low‐order finite volume (FV) methods applied to the v2 − f turbulence model of Durbin (Theoret. Comput. Fluid Dyn. 1991; 3:1–13) in the near vicinity of solid walls. We are not (like many others) concerned with the stability of solvers ‐ the topic at hand is simply whether the mathematical properties of the v2 − f model can be captured by the given, widespread, numerical method. The v2 − f model is integrated all the way up to solid walls, where steep gradients in turbulence parameters are observed. The full resolution of wall gradients imposes quite high demands on the numerical schemes and it is not evident that common (second order) FV codes can fully cope with such demands. The v2 − f model is studied in a statistically one‐dimensional, fully developed channel flow where we compare FV schemes with a highly accurate spectral element reference implementation. For the FV method a higher‐order face interpolation scheme, using Lagrange interpolation polynomials up to arbitrary order, is described. It is concluded that a regular second‐order FV scheme cannot give an accurate representation of all model parameters, independent of mesh density. To match the spectral element solution an extended source treatment (we use three‐point Gauss–Lobatto quadrature), as well as a higher‐order discretization of diffusion is required. Furthermore, it is found that the location of the first internal node need to be well within y+=1. Copyright © 2009 John Wiley & Sons, Ltd.
Relationship between white matter changes and cognition in healthy elders
Authors: Ota Miho , Nemoto Kiyotaka , Sato Noriko , Yamashita Fumio , Asada Takashi
Published: June 2009

Objectives

Cerebral WMHs on T2‐weighted magnetic resonance imaging (MRI) are common incidental findings in cognitively healthy elderly subjects. The relationship between such changes and cognitive function remains unclear.

Methods

The present study evaluated the relationship between the degree of white matter changes and cognitive function using data from 172 cognitively healthy subjects who underwent MRI and a battery of neuropsychological tests. The degree of WMHs was rated using a four‐point scale for images on a computer screen.

Results

Regarding the frontal and parieto‐occipital regions and basal ganglia region, compared with the group with no WMHs, the group with the most severe WMHs showed significantly lower performances for attention and disorientation to time, respectively.

Conclusions

Our results suggest that even in cognitively healthy elderly individuals, presence of large WMHs affects performance on certain cognitive domains according to their localization. Copyright © 2009 John Wiley & Sons, Ltd.
Circadian patterns of ad libitum smoking by menstrual phase
Authors: Allen Alicia M. , Mooney Marc , Chakraborty Rima , Allen Sharon S.
Published: June 2009

Objective

Recent research suggests nicotine metabolism may be influenced by sex hormones. Thus, we hypothesized that circadian smoking patterns would vary by menstrual phase.

Methods

Healthy female smokers (n = 31) between the ages of 18 and 40 with regular menstrual cycles, and not using hormones or psychotropic medications, were recruited for a randomized clinical study. Subjects recorded the time of each cigarette smoked and their menstrual phase with daily diaries prospectively for one complete menstrual cycle of ad libitum smoking. Analyses included Poisson regression to assess variations in the rate of smoking during waking hours (i.e., 6:00 a.m. and 12:00 midnight) and circadian smoking patterns by menstrual phase.

Results

Participants were 29.61 ± 5.44 years of age and smoked 16.93 ± 5.37 cigarettes per day. Participants had a lower rate of smoking during waking hours in the follicular phase as compared to the menses phase. There were no significant menstrual phase differences in the circadian smoking patterns.

Conclusions

These results offer further support for the influence of sex hormones on smoking behavior, but not on circadian patterns of smoking. Additional research is needed to study the direct relationship between nicotine metabolism, sex hormones, menstrual phase, and smoking behavior. Copyright © 2009 John Wiley & Sons, Ltd.
Enhancement of monoclonal antibody productivity by promoting active hypothermic growth in hybridoma cells
Authors: Chong Sun Li , Mou Duen Gang , Lim Saw Hoon , Ali Abdul Manaf , Tey Beng Ti
Published: June 2009

BACKGROUND: Many reports have suggested that mild hypothermic culture conditions improve the specific monoclonal antibody (mAb) productivity of mammalian cells. The effect of active hypothermic growth on the mAb productivity of the hybridoma C2E7 was investigated. Hybridoma growth under hypothermic conditions (32 °C) was stimulated by supplementation of the culture medium with high serum concentrations (up to 30%).

RESULTS: Specific and volumetric mAb productivity of a stimulated, active growth, mildly hypothermic hybridoma culture (30% FBS supplemented, 32 °C) were 1.38‐ and 1.34‐fold greater than the control culture (10% FBS supplemented, 37 °C). The enhanced specific mAb productivity under hypothermic conditions was associated with an increase in IgM mRNA levels during both the lag and early exponential phases of hypothermic growth.

CONCLUSION: Stimulation of hybridoma growth under mildly hypothermic conditions increased both the specific and volumetric mAb productivity of hybridoma cells. Copyright © 2009 Society of Chemical Industry
An evaluation of high resolution precipitation products at low resolution
Authors: Sapiano Mathew Raymond Paul
Published: June 2009

The large‐scale homogeneity of several high‐resolution satellite‐based estimates of precipitation is assessed against the Global Precipitation Climatology Project (GPCP) between January 2003 and December 2006. The high‐resolution estimates are aggregated to match the resolution of GPCP (monthly, 2.5°) and an Empirical Orthogonal Functions (EOF) analysis is conducted on each product along with an analysis of the slope from a linear regression with time. The results show that some of the datasets have significant artefacts and that none of the high‐resolution products give a good depiction of precipitation above 50°N. The two datasets which are most consistent with GPCP are the Tropical Rainfall Monitoring Mission (TRMM) Multi‐satellite Precipitation Analysis (TMPA) and the Global Satellite Mapping Project (GSMaP) both of which have undergone recent re‐processing which is an important quality for dataset homogeneity. Copyright © 2009 Royal Meteorological Society
Novel pH‐ and Temperature‐Responsive Methacrylamide Microgels
Authors: Pérez‐Álvarez Leyre , Sáez‐Martínez Virginia , Hernáez Estíbaliz , Katime Issa
Published: June 2009

A novel transition temperature in MeAM copolymer microgels is reported. Despite the fact that MeAM homopolymers do not show thermosensitive properties, a specific synthetic strategy leads to a thermo‐responsive swelling behavior that could be potentially useful in medical and/or industrial applications. The pH and temperature‐dependent swelling response of microgels of MeAM copolymerized with 2‐aminomethylpyridine and ethylenediamine is reported. The changes in particle sizes, which depend on the nature of the surrounding environment, are recorded by QELS. The relation between copolymer structure and its novel behavior is analyzed by several techniques (1H NMR, TGA).

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Taking the distortion of component parts along a manufacturing chain into consideration during planning
Authors: Klein D. , Seifert M. , Thoben K.‐D.
Published: June 2009

Distortion of a component part can already be minimized if it is considered in the planning of the manufacturing process. Thus, costs of the removal of distortion, e. g. by grinding, can be reduced. Beginning with steel production the state of a workpiece changes every process step continuously. Thereby, a distortion potential arises that will be visible at the component part after hardening as distortion. To consider distortion in planning the state has to be determined at every time of manufacturing. So far, no tools exist to describe the workpiece state along an entire process chain. In this article an alternative will be presented which is able to describe changes of states along an entire manufacturing chain. Finally, with this description an estimation of the expected distortion can be carried out.
Characterization of segregations and microstructure and their influence on distortion of low alloy SAE 5120 steel
Authors: Prinz C. , Hunkel M. , Clausen B. , Hoffmann F. , Zoch H.‐W.
Published: June 2009

Investigations within the SFB 570 “Distortion Engineering” showed that shafts produced of the low alloy steel SAE 5120 (EN 20MnCr5) changed curvature into a preferred direction after blank hardening. The changes of curvature could be correlated with a macroscopic structure, which was visible in the metallographic cross section of the hardened shafts. Due to a quantitative description of the orientation of the square structure within the metallographic cross section it could be shown that the changes of the orientation of the structure were similar to the changes of the preferred direction of the curvature of the blank hardened shafts.

Metallographic structures in different sections became visible because of differences of the local corrosion behavior. Such differences cause in varieties of the local chemical composition or grain size. Therefore the local chemical analysis was characterized by means of EPMA measurements with highest resolution. Further the grain size was described by quantitative image analysis of the former austenite grain boundaries. Additionally, characteristic areas of the metallographic cross section were investigated by FEM measurements due to the fact that the grain size could be affected by elemental precipitations.

The laminar EPMA‐measurements (mapping) at blank hardened shafts of the element distributions concerning the substantial alloying elements C, Mn and Cr showed a square structure in the element distribution. Best reproduction of the square structure within the metallographic cross section could be reached by moving averages of the highly resoluted mappings and their standard deviations.

Different distributions of the former austenite grain boundary were found within characteristic areas of the metallographic cross section. FEM measurements seem to point on the fact that differences of the former austenite grain size distributions could cause in varieties of the character and distribution of elemental precipitations.
Material flow analysis for hot‐forming of 20MnCr5 gear wheel blanks
Authors: Rentsch R.
Published: June 2009

During the manufacture of disk‐like gearwheel blanks made of 20MnCr6 (SAE 5120) a characteristic distortion behavior was observed which has been related to the forging process. The steel showed a typical banded structure and a distribution of chemical elements (segregations) perpendicular to rolling direction. When analyzing the micrographs of the forged gearwheel blanks, a significant variation in the material flow was found. In order to clarify the cause of the observed material flow variations and to determine its impact on the distortion potential of the disks, a twofold approach has been applied: a) to determine and to describe the material flow characteristics on basis of micrographs of forged disks and b) to carry out FE‐analyses on the theoretical material flow and its parameters. In this contribution results of the material flow analysis are presented and compared with FE‐analyses of the forming process.