bluuebunny/crossref_metadata_2025 huggingface dataset:
Simultaneous Determination of Aluminum and Iron in High Salt Content Matrices by Adsorptive Stripping Voltammetry. Application to Dialysis Fluids
Authors: Locatelli Clinio
Published: September 2003

A new analytical procedure for the simultaneous determination of aluminum(III) and iron(II) in two kinds of dialysis fluids (peritoneal and hemodialysis fluids) by differential pulse adsorptive stripping voltammetry (DPAdSV) is described. The voltammetric measurements were performed using, as working electrode, a stationary mercury electrode, and a platinum electrode and a Ag|AgCl|KCl(sat.) electrode as auxiliary and reference electrodes, respectively, employing acetate buffer solutions at different pH as supporting electrolyte. As complexing agents, Solochrome Violet RS, Palatine Chrome Black 6BN, Chromazurol S and Eriochrome Black T were employed. For both elements, the accuracy, expressed as relative recovery R%, was very satisfactory being in the range 94–105%, the precision as repeatability, expressed as relative standard deviation sr%, was lower than 6%, while the limits of detection were of the order of a few units of μg/L. The analytical voltammetric procedure has been validated by comparison with spectroscopic (graphite furnace atomic absorption spectroscopy, GFAAS) measurements.
Preparation of Fluorine‐Doped Lead Dioxide Modified Electrodes for Electroanalytical Applications
Authors: Ai Shiyun , Gao Mengnan , Zhang Wen , Sun Zhengdong , Jin Litong
Published: September 2003

The effect of F− on the modified films of lead dioxide in morphology and structure was studied. The results obtained by cyclic voltammetry (CV), X‐diffractometer (XRD) and scanning electron microscope (SEM) techniques indicated that F− could change the magnitude of lead dioxide crystal grain and the preferred crystallizing orientation on the substrate surface, even though it didn't change the basic structure of PbO2. When the modified electrode was applied as an analytical sensor to determine phenolic compounds, the linearity was in the range of 2×10−5 – 1×10−3 mol/L and the detection limit was 2.5×10−6 mol/L. It was all found that the stability and reproducibility of the oxide‐modified electrodes were improved by additional F−.
Solid Microelectrodes for In Situ Voltammetric Measurements
Authors: Pižeta Ivanka , Billon Gabriel , Fischer Jean‐Claude , Wartel Michel
Published: September 2003

Solid Au and Ag microelectrodes for both, model, and in situ voltammetric measurements, have been designed and tested. Amalgamation procedure and the parameters of measurement procedure are discussed. Differences between gold and silver electrodes were studied. Silver electrodes showed more stable behavior and were used prevalently for calibration and in situ measurements. Calibration of a 30‐μm diameter Ag electrode for Mn(II), Fe(II), I(−I) and S(−II) showed good linearity with reasonable confidence intervals, necessary for quantification of the signals obtained in situ, with average sensitivity of 0.05, 0.01, 0.16 and 1.10 nA/(μmol L−1) and minimal detection limit of 3.5, 28, 0.3 and 0.05 μmol L−1, respectively. Additional checking of the electrode sensitivity with the reduction peaks of oxygen is proposed.
Solution Equilibria. A Unified Mathematical Treatment of Experimental Polarographic and Potentiometric Data from AcidBase and Ligand Titrations. A Polarographic and Ion Selective Electrode Study of CdII(N‐(2‐hydroxyethyl)iminodiacetic acid)OH System
Authors: Cukrowski Ignacy , Maseko Nontombi
Published: September 2003

It has been demonstrated that potentiometric and polarographic data coming from either acidbase or ligand titrations can be evaluated by the same set of equations and mathematical procedures involving mass‐balance equations written for any metalligand model, including polynuclear species. It is shown that the concept of the complex formation curves, used previously in modeling and refinement of stability constants in acid‐base titration, is of general nature and can be successfully used in polarographic and potentiometric experiments conducted as a function of pH or an excess of a ligand. It appears that the linear relationship ΔE vs. log [M], typical for potentiometric studies with ISE, holds also in the case of the study of kinetically mixed metal‐ligand system studied by polarography when the corrected shift in the polarographic signal is used. The relationship ΔE vs. log [M] applies equally to the acid‐base and ligand titration for both experimental techniques employed (potentiometry and polarography). The significance of the corrected shift is discussed and its meaning in the study of kinetically fast or slow metalligand systems is elaborated. Advantages of the acid‐base titration over the ligand titration are discussed. The generalized mathematical data treatment was successfully employed in the study of the CdII(N‐(2‐hydroxyethyl)iminodiacetic acid) system. Results obtained from both analytical techniques (potentiometry with the use of an ion selective electrode, and two polarographic techniques) and analytical procedures (acidbase and ligand titrations) compare well with each other and with the literature data (the formation and stability constants of ML and ML2). In addition, a new complex M(HL) was identified and its stability constant is reported.
Alternating Current Tensammetry of Tetraphenylborate (TPB) and its Application for Potassium Determination
Authors: Ion Ana , Costache Madalina Beatrice , Grecu I. V. , Banica F. G.
Published: September 2003

Alternating‐current polarography of tetraphenylborate ion (TPB) in acetate buffer (pH 4 to 5) or 0.1 to 0.5 M NaCl produces a characteristic tensammetric (pseudocapacitive) peak at −0.95 V (vs. SCE). Within a limited range (till about 0.2 mM), the tensammetric peak current is proportional to the TPB concentration but it levels off at higher concentration. TPB concentration can also be determined within 5 to 50 μM by measuring the capacity current at −0.4 V or the differential pulse polarographic peak current at −0.85 V. The tensammetric peak current was used to determine the solubility of potassium tetraphenylborate in acetate buffer and sodium chloride solutions. Also, a method for amperometric back‐titration of potassium ion was worked out and applied to the analysis of mineral water.
Characteristics of Subtractive Anodic Stripping Voltammetry of Lead, Cadmium and Thallium at Silver‐Gold Alloy Electrodes
Authors: Bonfil Y. , Brand M. , Kirowa‐Eisner E.
Published: September 2003

Silver‐gold alloy electrodes have been studied for the purpose of the quantitative determination of heavy metals by subtractive anodic stripping voltammetry, (SASV). The results have been compared with those obtained with the silver and gold electrodes. The 50/50 a/o Ag/Au alloy electrode is the most suitable for quantifying thallium in the presence of lead and cadmium. The separation of its peak from those of lead and cadmium is 200 mV, which is about twice the separation obtained on the pure metal electrodes and is also better than on mercury. The silver electrode is suitable for the simultaneous determination of thallium, lead and cadmium. The peaks of lead and cadmium overlap on the 50/50 alloy. Pure silver or pure gold can be used for simultaneous quantification of these two metals. The use of gold for quantifying lead and cadmium is more limited because the peak potential of cadmium is shifted in the negative direction as its concentration increases and at [Cd2+]>200 nM, the two peaks merge. SASV enables correction for background currents and is of utmost importance for obtaining well‐defined peaks. The peaks of lead, cadmium and thallium appear over a relatively narrow potential range (ca. 200 mV) on all the electrodes presented in this work. For this reason, the quantifying of a peak is based on the derivative at the inflection point of only one of its branches (ascending or descending). All SASV measurements were carried out without removal of oxygen.
Using the Aggregation of Latex Polymers in the Fabrication of Reproducible Enzyme Electrodes
Authors: Rahmat Wibowo , Fischer Joshua , Hibbert D. Brynn , Gilbert Robert G. , Gooding J. Justin
Published: September 2003

An enzyme electrode for glucose is described as a model system to demonstrate a fabrication method using latex aggregation and entrapment of enzyme. Electrosterically‐stabilized latex particles synthesized by emulsion polymerization in batch from acrylic acid, methyl methacrylate and butyl acrylate, and glucose oxidase were coagulated together at pH 5.5 with ethanol. A platinum disk electrode dipped in the solution becomes coated with latex/enzyme. The relative thickness of the film and relative amount of enzyme may be controlled by the time the electrode is in contact with the solution. The enzyme was then immobilized by covalent attachment of amine groups to carboxylic moieties in the polymer using 1‐ethyl‐3(3‐dimethylaminopropyl)‐carbodiimide hydrochloride and N‐hydroxysuccinimide. Five minutes contact with the latex/enzyme solution and subsequent amide coupling, gave electrodes with a reproducibility of 5.7% RSD, a wide dynamic range (0–100 mM) and good storage properties.
Electroanalysis at Diamond‐Like and Doped‐Diamond Electrodes
Authors: Compton Richard G. , Foord John S. , Marken Frank
Published: September 2003

Diamond as a high performance material occupies a special place due to its in many ways extreme properties, e.g., hardness, chemical inertness, thermal conductivity, optical properties, and electric characteristics. Work mainly over the last decade has shown that diamond also occupies a special place as an electrode material with interesting applications in electroanalysis. When made sufficiently electrically conducting for example by boron‐doping, ‘thin film' and ‘free–standing' diamond electrodes exhibit remarkable chemical resistance to etching, a wide potential window, low background current responses, mechanical stability towards ultrasound induced interfacial cavitation, a low ‘stickiness' in adsorption processes, and a high degree of ‘tunability' of the surface properties. This review summarizes some of the recent work aimed at applying conductive (boron‐doped) diamond electrodes to improve procedures in electroanalysis.
Quantification of analytes in overlapping peaks from capillary electrophoresis using multivariate curve resolution‐alternating least squares methods
Authors: Li Hua , Zhang Fang , Havel Josef
Published: September 2003

Application of multivariate curve resolution with alternating least squares (ALS) methods to second‐order data from capillary electrophoresis diode array detector (CE‐DAD) is shown. Second‐order data are easily obtained by setting individual data matrix of CE run one in top of the other. Initial qualitative solutions obtained by evolving factor analysis can be further optimized by simultaneous analysis of multiple electrophoresis run data with ALS regression. Quantification is achieved by the comparison of the analyte peak areas with that of pure standards. During the ALS regression procedure, the following constraints were applied: (i) both concentrations and unit pure spectra of the resolved components must be positive; (ii) elution profiles have an unimodal shape; (iii) correspondence exists between common species in the different data matrices; (iv) the pure spectrum of each species is the same in all runs where it is present. The above methods were applied for the determination of dinitrotoluene (DNT) isomeric compounds in overlapping peaks from CE.
Monitoring of chemical reactions within microreactors using an inverted Raman microscopic spectrometer
Authors: Fletcher Paul D. I. , Haswell Stephen J. , Zhang Xunli
Published: September 2003

An inverted Raman microscope spectrometer has been used to profile the spatial evolution of reactant and product concentrations for a chemical reaction within a microreactor operating under hydrodynamic flow control. The Raman spectrometer was equipped with a laser source at wavelength of 780 nm, confocal optics, a holographic transmission grating, and a charge‐coupled device (CCD) detector. The microreactor consisted of a T‐shaped channel network etched within a 0.5 mm thick glass bottom plate that was thermally bonded to a 0.5 mm thick glass top plate. The ends of the channel network were connected to reagent reservoirs that were linked to a syringe pump for driving the solutions by hydrodynamic pumping within the channels. The microchannels were 221 μm wide and 73 μm deep. The synthesis of ethyl acetate from ethanol and acetic acid was investigated as a model system within the microreactor as Raman scattering bands for each reactant and product species were clearly resolved. Raman spectral intensities of each band were proportional to concentration for each species and hence all concentrations could be quantitatively measured after calibration. By scanning specific Raman bands within a selected area in the microchannel network at given steps in the X‐Y plane, spatially resolved concentration profiles were obtained under steady‐state flow conditions. Under the flow conditions used, different positions within the concentration profile correspond to different times after contact and mixing of the reagents, thereby enabling one to observe the time dependence of the product formation. Raman microscopy provides a useful complementary technique to UV/VIS absorbance and fluorescence methods for the in situ monitoring and analysis of chemical reaction species having their lowest S0‐S1 absorption bands too far in the UV to be of use, due to their probable overlap with the bands from other reactant, product and solvent molecules.
Direct tris(2,2'‐bipyridyl)ruthenium (II) electrochemiluminescence detection of polyamines separated by capillary electrophoresis
Authors: Liu Jifeng , Yang Xiurong , Wang Erkang
Published: September 2003

Capillary electrophoresis (CE) with tris(2,2`‐bipyridyl) ruthenium (II) (Ru(bpy)32+) electrochemiluminescence (ECL) detection technique was developed for the analysis of four polyamines (putrescine (Put), cadaverine (Cad), spermidine (Spd), and spermine (Spm)) analysis. The four polyamines contain different amine groups, which have different ECL activity. There are several parameters which influence the resolution and ECL peak intensities, including the buffer pH and concentrations, separation voltage, sample injection, electrode materials, and Ru(bpy)32+ concentrations. Polyamines are separated by capillary zone electrophoresis in an uncoated fused‐silica capillary (50 cm×25 μm (ID) filled with acidic phosphate buffer (200 mmol/L phosphate, pH 2.0) – 1mol/L phosphoric acid (9:1 v/v) and a separation voltage of 5 kV (25 μA), with end‐column Ru(bpy)32+ ECL detection. A 5 mmol/L Ru(bpy)32+ solution plus 200 mmol/L phosphate buffer (pH 11.0) is added into the reagent reservoir. The calibration curve is linear over a concentration range of two or three orders of magnitude for the polyamines. The analysis time is less than 25 min. Detection limits for Put and Cad are 1.9×10−7 mol/L and 7.6×10−9 mol/L for Spd and Spm, respectively. Intraday and interday relative standard deviations of ECL peak intensities are less than 8%. The main advantages of this CE‐ECL detection technique for polyamines analysis presented herein are the omission of chemical derivatization of the analytes and the high selectivity.
Influence of pH on electrophoretic behavior of phenothiazines and determination of pKa values by capillary zone electrophoresis
Authors: Lin Ching‐Erh , Liao Wei‐Ssu , Chen Kuo‐Hsing , Lin Wann‐Yin
Published: September 2003

The influence of buffer pH on the electrophoretic behavior of 13 structurally related phenothiazines and determination of pKa values by capillary zone electrophoresis (CZE) were investigated. The results indicate that phenothiazines with a piperazine substituent behave quite differently from those with substituents having an aliphatic side chain or a piperidine moiety over the pH range studied. To separate these phenothiazines, it is preferable to select buffer pH in the range of 2.5–3.5. The pKa values of phenothiazines with three different types of substituents attached at the 10‐position of the phenothiazine ring were determined. The determination of pKa values of phenothiazines allows us to rationalize the influence of buffer pH on the migration behavior of these compounds in CZE.
Comprehensive two‐dimensional separation system by coupling capillary reverse‐phase liquid chromatography to capillary isoelectric focusing for peptide and protein mapping with laser‐induced fluorescence detection
Authors: Mao Yu , Zhang Xiangmin
Published: September 2003

A comprehensive two‐dimensional (2‐D) separation system, coupling capillary reverse‐phase liquid chromatography (cRPLC) to capillary isoelectric focusing (CIEF), is described for protein and peptide mapping. cRPLC, the first dimension, provided high‐resolution separations for salt‐free proteins. CIEF, the second dimension with an orthogonal mechanism to cRPLC afforded excellent resolution capability for proteins with efficient protein enrichment. Since all sample fractions in cRPLC effluents could be transferred to the CIEF dimensions, the combination of the two high‐efficiency separations resulted in maximal separation capabilities of each dimension. Separation effectiveness of this approach was demonstrated using complex protein/peptide samples, such as yeast cytosol and a BSA tryptic digest. A peak capacity of more than 10 000 had been achieved. A laser‐induced fluorescence (LIF) detector, developed for this system, allowed for high‐sensitive detection, with a fmol level of peptide detection for the BSA digest. FITC and BODIPY maleimide were used to tag the proteins, and the latter was found better both for separation and detection in our 2‐D system.
Separation and determination synchronously by multichannel mode‐filtered light capillary electrochromatography
Authors: Zuo Xinbing , Wang Kemin , Zhou Leiji , Huang Shasheng
Published: September 2003

Mode‐filtered light capillary electrophoresis (CE) was developed for capillary electrochromatography (CEC) by applying annular columns of different size (250, 320, and 530 μm) and a bovine serum albumin (BSA)‐modified chiral stationary phase. BSA was encapsulated on the pretreated surface of an optic fiber and the inner wall of a capillary using sol‐gel technique with a resultant larger active stationary phase area. The coating was examined by atomic force microscopy (AFM) and Fourier transform infrared (FT‐IR). The separation of DL‐tryptophan (10 mM) was investigated under different voltage/current conditions.
Direct immunoassay of estrone by capillary electrophoresis with laser‐induced fluorescence detection
Authors: Su Ping , Zhang Xin‐Xiang , Chang Wen‐Bao
Published: September 2003

An immunoassay for estrone (E1) in women's serum, based on the competitive reaction between fluorescein‐labeled complete antigen and E1 with limited amount of anti‐estrone monoclonal antibody is described. A thermally reversible hydrogel, poly‐N‐isopropylacrylamide (pNIPA), was added to the buffer to improve the reproducibility. With a laser‐induced fluorescence (LIF) detector, the capillary electrophoretic immunoassay (CEIA) can be applied to determine E1 at a concentration lower than 19.6 pg/mL. The E1 levels in ten normal women's serum were measured at the range of 118.6–222.0 pg/mL.
Critical factors for reproducible capillary electrophoresis of Escherichia coli cells
Authors: Dai Dongsheng , Chen Yi , Qi Li , Yu Xiao
Published: September 2003

Sharp peaks of Escherichia coli JM 109 (up to 1 300 000 theoretical plates) were recorded with either extremely diluted (< 5 mM) or extremely concentrated (ca. 150 mM) Tris‐borate (TB) running buffers. However, under the conditions of yielding sharp peaks, migration time of E. coli was irreproducible. Critial factors influencing reproducibility were found to include bacterial growth phase, storage condition, cell pretreatment before injection, and concentration of running buffer. Buffer concentrations in the range of 20–100 mM TB were essential for reproducibility. E. coli JM 109 was shown to be sensitive to ultrasonification. Bacterial growth and storage conditions could be monitored by CE, with results comparable to those obtained with optical methods.
Direct computation of thermodynamic properties of chemically reacting air with consideration to CFD
Authors: Iannelli Joe
Published: September 2003

This paper details a two‐equation procedure to calculate exactly mass and mole fractions, pressure, temperature, specific heats, speed of sound and the thermodynamic and jacobian partial derivatives of pressure and temperature for a five‐species chemically reacting equilibrium air. The procedure generates these thermodynamic properties using as independent variables either pressure and temperature or density and internal energy, for CFD applications. An original element in this procedure consists in the exact physically meaningful solution of the mass‐fraction and mass‐action equations. Air‐equivalent molecular masses for oxygen and nitrogen are then developed to account, within a mixture of only oxygen and nitrogen, for the presence of carbon dioxide, argon and the other noble gases within atmospheric air. The mathematical formulation also introduces a versatile system non‐dimensionalization that makes the procedure uniformly applicable to flows ranging from shock‐tube flows with zero initial velocity to aerothermodynamic flows with supersonic/hypersonic free‐stream Mach numbers. Over a temperature range of more than 10000 K and pressure and density ranges corresponding to an increase in altitude in standard atmosphere of 30000 m above sea level, the predicted distributions of mole fractions, constant‐volume specific heat, and speed of sound for the model five species agree with independently published results, and all the calculated thermodynamic properties, including their partial derivatives, remain continuous, smooth, and physically meaningful. Copyright © 2003 John Wiley & Sons, Ltd.
Molecular analysis of t(X;11)(q24;q23) in an infant with AML‐M4
Authors: Fu Jen‐Fen , Liang Der‐Cherng , Yang Chao‐Ping , Hsu Jia‐Jong , Shih Lee‐Yung
Published: September 2003

t(X;11)(q24;q23) is a recurring chromosomal translocation in pediatric acute myeloid leukemia. The rearrangement results in fusion of MLL at 11q23 with SEPT6 at Xq24. Here, we report the identification of an MLL‐SEPT6 fusion transcript in an infant with acute myeloid leukemia (AML)‐M4. Reverse transcription–polymerase chain reaction confirmed the presence of an MLL‐SEPT6 fusion transcript composed of exon 8 of MLL and exon 2 of SEPT6, but the absence of the reciprocal SEPT6‐MLL fusion transcript. Sequence analysis of the genomic break junctions in MLL and SEPT6 suggested that the rearrangement in this case was the result of an insertion of the inverted Xq24 segment, which contained the 3′ region of SEPT6 intron 1 and up to 950 kb centromeric to SEPT6, into MLL intron 8. This is a novel type of chromosomal rearrangement leading to the MLL‐SEPT6 fusion. The presence of deletions, duplications, and non‐template DNA sequence at the break junctions suggested that the DNA damage‐repair machinery is likely to be involved in the translocation events. © 2003 Wiley‐Liss, Inc.
High‐resolution mapping of amplifications and deletions in pediatric osteosarcoma by use of CGH analysis of cDNA microarrays
Authors: Squire Jeremy A. , Pei Jianming , Marrano Paula , Beheshti Ben , Bayani Jane , Lim Gloria , Moldovan Laura , Zielenska Maria
Published: September 2003

Conventional cytogenetic and comparative genomic hybridization (CGH) studies have shown that osteosarcomas (OSs) are characterized by complex structural and numerical chromosomal alterations and gene amplification. In this study, we used high‐resolution CGH to investigate recurrent patterns of genomic imbalance by use of DNA derived from nine OS tumors hybridized to a 19,200‐clone cDNA microarray. In six OSs, there was copy number gain or amplification of 6p, with a minimal region of gain centering on segment 6p12.1. In seven OSs, the pattern of amplification affecting chromosome arm 8q showed high‐level gains of 8q12–21.3 and 8q22–q23, with amplification of the MYC oncogene at 8q24.2. Seven OSs showed copy number gain or amplification of 17p between the loci bounded by GAS7 and PMI (17p11.2–17p12), and three of these tumors also showed small losses at 17p13, including the region containing TP53. An in silico analysis of the distribution of segmental duplications (duplicons) in this region identified a large number of tracts consisting of paralogous sequences mapping to the 17p region, encompassing the region of deletions and amplifications in OS. Interestingly, within this same region there were clusters of duplicons and several genes that are expressed during bone morphogenesis and in OS. In summary, microarray CGH analysis of the chromosomal imbalances of OS confirm the overall pattern observed by use of metaphase CGH and provides a more precise refinement of the boundaries of genomic gains and losses that characterize this tumor. © 2003 Wiley‐Liss, Inc.
Molecular cytogenetic characterization of the KG‐1 and KG‐1a acute myeloid leukemia cell lines by use of spectral karyotyping and fluorescence in situ hybridization
Authors: Mrózek Krzysztof , Tanner Stephan M. , Heinonen Kristiina , Bloomfield Clara D.
Published: September 2003

The KG‐1 cell line, established from bone marrow cells of a patient with erythroleukemia evolving to acute myelogenous leukemia, and its less differentiated variant, KG‐1a, are widely used in research worldwide. However, to our knowledge, neither cell line was studied by use of molecular‐cytogenetic techniques such as spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH). Our G‐banding, SKY, and FISH analyses revealed a complex karyotype with a pseudodiploid modal chromosome number in both the KG‐1 and KG‐1a cell lines. The lines shared several identical structural aberrations, including der(4)t(4;8), del(7q), der(8)t(8;12), idic(8)(p11), der(17)t(5;17), and der(20)t(12;20), but also differed with regard to other chromosome rearrangements. In contrast to KG‐1, the KG‐1a line lost one of the two copies of idic(8)(p11) present in KG‐1 cells and gained a der(8;22)(q24;q13), an i(11)(q10), and a der(19)t(14;19)(q13;q13.4). Notably, we have shown that the KG‐1 cells harbor a partial hexasomy of the long arm of chromosome 8, which may explain in part the previously reported significantly higher rate of formation of the AML1‐ETO fusion gene in KG‐1 cells subjected to high‐dose gamma irradiation compared with the rates of formation of the BCR‐ABL or the DEK‐CAN fusion gene. Our detailed description of chromosome rearrangements in KG‐1 and KG‐1a will be useful for the cytogenetic authentication of the lines, and provide clues as to the regions of the genome that could be studied further to explain the differences in phenotypic properties between KG‐1 and KG‐1a cells. © 2003 Wiley‐Liss, Inc.
Further cytogenetic characterization of multiple myeloma confirms that 14q32 translocations are a very rare event in hyperdiploid cases
Authors: Smadja Nicole Véronique , Leroux Dominique , Soulier Jean , Dumont Sylvie , Arnould Catherine , Taviaux Sylvie , Taillemite Jean Louis , Bastard Christian
Published: September 2003

Translocations involving the immunoglobulin heavy‐chain genes are frequent in multiple myeloma (MM), which can be separated into two groups according to the chromosome number pattern. 14q32 translocations 14q32t are more frequent in hypodiploid than in hyperdiploid karyotypes. However, conventional cytogenetics (CC) misses cryptic translocations, especially t(4;14)(p16;q32). Furthermore, recent interphase fluorescence in situ hybridization (FISH) studies found 14q32t in as many as 75% of MM cases. To identify in which CC group we failed to detect translocations, we designed a study by use of FISH with a dual‐color IGH probe on previously R‐banded metaphase cells, allowing the detection of both 14q32t and overall chromosomal abnormalities, in a new series of 55 MM with abnormal karyotypes: 4/29 hyperdiploid (14%) and 19/26 hypodiploid (73%) cases had a 14q32t. The t(4;14) was found in 2 hyperdiploid (7%) and 10 hypodiploid (39%) cases. We therefore confirm that 14q32t are much more frequent in hypodiploid than in hyperdiploid MM (P < 0.0001) and that cryptic t(4;14)(p16;q32) is strongly associated with hypodiploid karyotypes (P < 0.01). Through the use of this reliable assay, only 42% of MM had 14q32t. © 2003 Wiley‐Liss, Inc.
Expression profiling targeting chromosomes for tumor classification and prediction of clinical behavior
Authors: Lu Yong‐Jie , Williamson Daniel , Wang Rubin , Summersgill Brenda , Rodriguez Sandrine , Rogers Simon , Pritchard‐Jones Kathy , Campbell Colin , Shipley Janet
Published: September 2003

Tumors are associated with altered or deregulated gene products that affect critical cellular functions. Here we assess the use of a global expression profiling technique that identifies chromosome regions corresponding to differential gene expression, termed comparative expressed sequence hybridization (CESH). CESH analysis was performed on a total of 104 tumors with a diagnosis of rhabdomyosarcoma, leiomyosarcoma, prostate cancer, and favorable‐histology Wilms tumors. Through the use of the chromosome regions identified as variables, support vector machine analysis was applied to assess classification potential, and feature selection (recursive feature elimination) was used to identify the best discriminatory regions. We demonstrate that the CESH profiles have characteristic patterns in tumor groups and were also able to distinguish subgroups of rhabdomyosarcoma. The overall CESH profiles in favorable‐histology Wilms tumors were found to correlate with subsequent clinical behavior. Classification by use of CESH profiles was shown to be similar in performance to previous microarray expression studies and highlighted regions for further investigation. We conclude that analysis of chromosomal expression profiles can group, subgroup, and even predict clinical behavior of tumors to a level of performance similar to that of microarray analysis. CESH is independent of selecting sequences for interrogation and is a simple, rapid, and widely accessible approach to identify clinically useful differential expression. © 2003 Wiley‐Liss, Inc.
Molecular cloning of a constitutional t(7;22) translocation associated with risk of hematological malignancy
Authors: Hill Alexander S. , MacCallum Peter K. , Young Bryan D. , Lillington Debra M.
Published: September 2003

We report the molecular characterization of a reciprocal constitutional translocation t(7;22)(p13;q11.2) carried by three family members who have each developed a hematological malignancy. The chromosome 7 breakpoint was localized to a single BAC clone, RP11‐571N3, by sequential fluorescence in situ hybridization analysis of clones selected from the NCBI chromosome 7 map. This was further refined to a 739‐bp region by Southern blot analysis of DNA from the two cell lines 1193 and 1194 digested with EcoRI, HindIII, PstI, and PvuII. A 2.8‐kb fragment spanning the der(22) breakpoint was amplified by long‐range inverse PCR. The sequence of this fragment was used to predict the composition of the der(7) breakpoint, and a 1.3‐kb fragment was amplified by use of primers from both chromosomes 7 and 22 based on this prediction. The breakpoint on chromosome 22 is located between the 3rd and 4th V regions of the immunoglobulin lambda (IGL) locus, and the breakpoint on chromosome 7 is located 122 kb proximal to the insulin‐like growth factor binding protein (IGFBP) 3 gene. Examination of both reciprocal junctions showed that four bases were lost from chromosome 22, whereas 75 bases were lost from chromosome 7. Small insertions of 46 bases and 13 bases were found at the der(22) and the der(7) junctions, respectively. As a consequence of this event, the entire IGL locus, less the first three Vλ elements, is translocated to chromosome 7, whereas the three remaining Vλ elements on the der(22) are juxtaposed with IGFBP3 and IGFBP1. © 2003 Wiley‐Liss, Inc.
Clustering of hypermethylated genes in neuroblastoma
Authors: van Noesel Max M. , van Bezouw Saskia , Voûte P. A. , Herman James G. , Pieters Rob , Versteeg Rogier
Published: September 2003

CpG‐island hypermethylation of gene promoters is a frequent mechanism for gene inactivation in tumors. Many neuroblastomas have hypermethylation and down‐regulation of CASP8, leading to resistance to tumor necrosis factor–related apoptosis‐inducing ligand (TRAIL). We recently found hypermethylation of the four TRAIL receptors in 9 neuroblastoma cell lines. Here, we analyzed methylation of 34 genes in 22 neuroblastoma cell lines. Of the 29 newly analyzed genes, only FLIP at chromosome band 2q33 was methylated in 8/22 cell lines. The FLIP protein is a negative regulator of Caspase 8. FLIP maps adjacent to CASP8, and their methylation patterns showed a moderate correlation. Furthermore, co‐methylation patterns were observed for the TRAIL receptor pairs DCR1 and DCR2 and between DR4 and DR5. All four receptors co‐localize in chromosome band 8p21. The 6 genes methylated in neuroblastomas appeared to occur in pairs. The genes within each pair have a strong sequence homology and originated from gene duplication. We found no evidence for regional spreading of methylation, given that we did not observe de novo methylation in additional local CpG islands. However, the gene pairs showed a striking co‐regulation at the mRNA expression level. Down‐regulation of FLIP strongly corresponds with down‐regulation of CASP8, and this was also found for DCR1 and DCR2. Only a subset of the down‐regulated genes was methylated. This suggests a mechanism of co‐regulated transcriptional silencing of the gene pairs, followed by a methylation event that is less penetrating. The methylation pattern therefore supports a model in which CpG islands are not randomly targeted by methylation in cancer. Specific transcriptional silencing probably marks genes that can become methylated. © 2003 Wiley‐Liss, Inc.
Aberrant expression of the LHX4 LIM‐homeobox gene caused by t(1;14)(q25;q32) in chronic myelogenous leukemia in biphenotypic blast crisis
Authors: Yamaguchi Mitsuko , Yamamoto Koh , Miura Osamu
Published: September 2003

Chromosomal aberrations observed in addition to the Philadelphia chromosome in chronic myelogenous leukemia (CML) are likely to be involved in disease progression to the blast crisis. We describe here a t(1;14)(q25;q32) as an additional chromosomal aberration in a patient with CML in biphenotypic blast crisis. By use of long‐distance inverse polymerase chain reaction (PCR), we cloned the chromosomal breakpoint and revealed that the immunoglobulin heavy chain gene is fused near its Eμ enhancer region to the 5′ region of the LHX4 LIM‐homeobox gene, whose expression is restricted to the central nervous system. By use of quantitative real‐time reverse‐transcription PCR, we found that the LHX4 mRNA is expressed at high levels in the patient's leukemic cells and in an acute lymphoblastic leukemia (ALL) cell line. The aberrant expression of the LHX4 gene by the t(1;14)(q25;q32) has very recently been reported in a case of ALL, thus, representing a rare, but recurrent genetic abnormality of possible importance in leukemogenesis. © 2003 Wiley‐Liss, Inc.
Development of a real‐time RT‐PCR assay for the quantification of the most frequent MLL/AF9 fusion types resulting from translocation t(9;11)(p22;q23) in acute myeloid leukemia
Authors: Scholl Claudia , Breitinger Heike , Schlenk Richard F. , Döhner Hartmut , Fröhling Stefan , Döhner Konstanze
Published: September 2003

One strategy to predict clinical outcome in patients with acute myeloid leukemia (AML) is detection of minimal residual disease (MRD) after achievement of hematologic complete remission (CR). We established a real‐time RT‐PCR assay by use of TaqMan technology for the identification of MRD by quantification of the most frequent fusion transcripts resulting from t(9;11)(p22;q23). To achieve comparable PCR efficiencies between the different PCR assays, primers were chosen to obtain amplicons of nearly identical lengths. MLL/AF9 copy numbers were normalized to the housekeeping gene porphobilinogen deaminase (PBGD). The sensitivity of the assay, as determined at the cellular level, was comparable to that of qualitative single‐round RT‐PCR. Samples from eight patients with t(9;11)‐positive AML were analyzed. At diagnosis and relapse, normalized copy numbers were positive and ranged from 490 to 5,558. Samples from two of seven patients collected at the time of CR became negative, whereas five cases still had positive normalized copy numbers with values between 5 and 5,286. The implications of MRD detection by MLL/AF9 fusion transcript quantification for the clinical management of t(9;11)‐positive AML have to be determined in further studies. © 2003 Wiley‐Liss, Inc.
Pattern of secondary genomic changes in pancreatic tumors of Tgfα/Trp53+/− transgenic mice
Authors: Schreiner Bettina , Baur Dorothee M. , Fingerle Alexander A. , Zechner Ulrich , Greten Florian R. , Adler Guido , Sipos Bence , Klöppel Günter , Hameister Horst , Schmid Roland M.
Published: September 2003

Trp53+/− mice overexpressing Tgfα in a pancreas‐specific manner represent a well‐established animal model for pancreatic cancer. In this study we analyzed 38 pancreatic adenocarcinomas of these mice for secondary genomic changes by comparative genomic hybridization (CGH), loss of heterozygosity (LOH) analysis, real‐time PCR, and methylation‐specific analysis. CGH screening of the tumors revealed a recurrent pattern of genomic changes. In more than 50% of the tumors, chromosome 11 was affected. The gain of the proximal part spans about 16 cM, including the genes for Egfr, Rel, and Stk10. The distal part of chromosome 11, which contains the Trp53 locus, was deleted. LOH analysis proved that almost all tumors segregate the wild‐type Trp53 allele. The Cdkn2a locus on chromosome 4 was inactivated by hypermethylation in 55% of all tumors. In addition, two other changes were detected in a mutually exclusive manner: overrepresentation of part of chromosome 15, or more rarely, loss of the distal part of chromosome 14. Together these data suggest the induction of a uniform pattern of secondary genomic changes in this transgenic tumor model for pancreatic cancer. © 2003 Wiley‐Liss, Inc.
CDKN2A as a uveal and cutaneous melanoma susceptibility gene
Authors: Kannengiesser Caroline , Avril Marie‐Françoise , Spatz Alain , Laud Karine , Lenoir Gilbert M. , Bressac‐de‐Paillerets Brigitte
Published: September 2003

A few families have been described whose members are affected by either cutaneous melanoma (CM) or uveal melanoma (UM), suggesting that a common susceptibility could exist. Although CDKN2A is the main CM predisposing gene, thus far no germline CDKN2A mutations have been described in families with both CM and UM. We report a Gly67Ser missense CDKN2A germline mutation in a melanoma‐prone family, where one carrier was affected by UM and the other by a CM. Immunohistochemistry performed on the UM tissue block revealed loss of CDKN2A protein staining in tumor cells. These observations demonstrate that CDKN2A is also a UM susceptibility gene. © 2003 Wiley‐Liss, Inc.
Compensatory cortical activation observed by fMRI during a cognitive task at the earliest stage of multiple sclerosis
Authors: Audoin Bertrand , Ibarrola Danielle , Ranjeva Jean‐Philippe , Confort‐Gouny Sylviane , Malikova Irina , Ali‐Chérif André , Pelletier Jean , Cozzone Patrick
Published: September 2003

Recent functional magnetic resonance imaging (fMRI) studies have suggested that functional cortical changes seen in patients with early relapsing‐remitting multiple sclerosis (MS) can have an adaptive role to limit the clinical impact of tissue injury. To determine whether cortical reorganization occurs during high cognitive processes at the earliest stage of multiple sclerosis (MS), we performed an fMRI experiment using the conventional Paced Auditory Serial Addition Test (PASAT) as paradigm in a population of ten patients with clinically isolated syndrome suggestive of multiple sclerosis (CISSMS). At the time of the fMRI exploration, mean disease duration was 6.8 ± 3.3 months. We compared these results to those obtained in a group of ten education‐, age‐, and sex‐matched healthy controls. Subjects were explored on a 1.5 T MRI system using single‐shot gradient‐echo EPI sequence. Performances of the two groups during PASAT recorded inside the MR scanner were not different. Statistical assessment of brain activation was based on the random effect analysis (between‐group analysis two‐sample t‐test P < 0.005 confirmed by individual analyses performed in the surviving regions P < 0.05 Mann Whitney U‐test). Compared to controls, patients showed significantly greater activation in the right frontopolar cortex, the bilateral lateral prefrontal cortices, and the right cerebellum. Healthy controls did not show greater activation compared to CISSMS patients. The present study argues in favor of the existence of compensatory cortical activations at the earliest stage of MS mainly located in regions involved in executive processing in patients performing PASAT. It also suggests that fMRI can evidence the active processes of neuroplasticity contributing to mask the clinical cognitive expression of brain pathology at the earliest stage of MS. Hum. Brain Mapping 20:51–58, 2003. © 2003 Wiley‐Liss, Inc.
How Does University Decision Making Shape the Faculty?
Authors: Cross John G. , Goldenberg Edie N.
Published: September 2003

The authors discuss important institutional changes that they view as probably permanennt for reasons of cost and that may have far‐reaching implications for the future of higher education.
Urinary IL‐2 assay for monitoring intravesical bacillus Calmette‐Guérin response of superficial bladder cancer during induction course and maintenance therapy
Authors: Saint Fabien , Kurth Nathalie , Maille Pascale , Vordos Dimitri , Hoznek Andre , Soyeux Pascale , Patard Jean Jacques , Abbou Claude C. , Chopin Dominique K.
Published: September 2003

We evaluated the clinical significance of Th1(IL‐2)/Th2(IL‐10) urinary profiles during a weekly induction course lasting 6 weeks, followed by a weekly maintenance therapy schedule for 3 weeks. Urinary IL‐2 and /IL‐10 were measured by ELISA in 39 patients receiving BCG for superficial bladder cancer or carcinoma in situ. Measurements were made after each instillation of 81 mg of BCG Connaught (Immucyst) during the induction course and the 3‐week maintenance therapy (given at 3, 6, 12, 18, 24, 30 and 36 months). Cytokine levels were correlated with the risk of recurrence, progression, leukocyturia and adverse events. Median follow‐up was 35 months (range 7–72 months). Complete responses to BCG were obtained in 30 patients (77%); the remaining 9 patients relapsed (23%), and 4 of these patients progressed (10.2%). Failure to detect urinary IL‐2 during BCG induction course and the first extended induction cycle (6+3 schedule) correlated with time to recurrence (p = 0.01) and progression (p = 0.01). During the extended induction cycle, the first instillation was associated with an IL‐2 cytokine profile, whereas the second and third instillations were associated with a switch to an IL‐10 cytokine profile. This switch was associated with leukocyturia (p = 0.0001) and adverse events (p = 0.03). The 6+3 schedule is associated with urinary IL‐2 overproduction and improved recurrence‐ and progression‐free survival. During the BCG extended induction cycle, the favorable IL‐2 urinary cytokine pattern gradually switches to an IL‐10 profile, suggesting that the schedule based on 3 weekly instillations may be unsuitable for some patients and that the dose and frequency of maintenance BCG instillations may be adapted to individual urinary cytokine levels. © 2003 Wiley‐Liss, Inc.
Tobacco smoking and chewing, alcohol drinking and lung cancer risk among men in southern India
Authors: Gajalakshmi Vendhan , Hung Rayjean J. , Mathew Aleyamma , Varghese Cherian , Brennan Paul , Boffetta Paolo
Published: September 2003

In India, lung cancer is one of the most common and lethal cancers, and tobacco smoking remains its most important etiologic factors. The objective of our study is to examine the effects of different tobacco consumption forms, including smoking and chewing, on lung cancer risk of men in southern India, especially to compare the effects of bidi smoking to cigarette smoking on lung carcinogenesis. We also evaluated the possible role of Indian alcohol beverages and non‐Indian alcohol beverages on lung carcinogenesis. We conducted a case‐control study in Chennai and Trivandrum. In total, 778 lung cancer cases and 3,430 controls, including 1,503 cancer controls and 1,927 healthy controls, were recruited. The effects of cigarette, bidi smoking, chewing and alcohol drinking on the risk of lung cancer were estimated from unconditional multivariate logistic regression. We also applied the generalized additive model (GAM) with locally‐weighted running‐line smoothers (loess) to find the most plausible curve for the dose‐response relationship. The results from GAM suggest a plateau after 35 years of smoking or 10 cigarette‐equivalent pack‐years for both cigarette and bidi. The OR is 4.54 (95%CI=2.96–6.95) and 6.45 (95%CI=4.38–9.50) for more than 30 years of cigarette‐only and bidi‐only smoking, respectively, and 6.87 (95%CI=4.62–10.2) and 10.7 (95%CI=5.82–19.6) for more than 12 weighted cumulative cigarette‐only and bidi‐only consumption, respectively. The lung cancer risk of former cigarette smokers drops down more quickly after quitting smoking compared to former bidi smokers. There is no evidence for the effect of chewing and lung cancer risk nor clear evidence of an effect of overall alcohol drinking among never‐smokers, although Indian alcohol drinking seemed to remain associated with lung cancer risk under limited power (OR=2.67, 95%CI=1.02–7.02). Bidi smoking seems to have a stronger carcinogenic effect than cigarette smoking: this difference holds no matter which aspect of smoking was considered. © 2003 Wiley‐Liss, Inc.
Infrequent microsatellite instability in liver fluke infection‐associated intrahepatic cholangiocarcinomas from Thailand
Authors: Liengswangwong Upama , Nitta Takayuki , Kashiwagi Hironobu , Kikukawa Hiroko , Kawamoto Toru , Todoroki Takeshi , Uchida Kazuhiko , Khuhaprema Thiravud , Karalak Anant , Srivatanakul Petcharin , Miwa Masanao
Published: September 2003

The liver fluke infection‐associated intrahepatic cholangiocarcinoma (ICC) is a major liver cancer in Northeast Thailand. The molecular basis of this ICC is poorly understood. To address possible roles of the DNA mismatch repair (MMR) system in ICC carcinogenesis, a fluorescence‐labeling PCR/laser scanning technique with high sensitivity was employed to analyze genomic instability in the nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) in 24 fresh and 13 formalin‐fixed, paraffin‐embedded tissues of ICC and their corresponding normal parts. Microsatellite instability (MSI) was assessed in nDNA, using 12 highly polymorphic loci including 5 Bethesda markers. These loci were mainly related to major MMR genes, hMSH2 and hMLH1. Also 3 (C)n and/or (C)n(A)n repeat instability at 1 noncoding region in the displacement‐loop (D‐loop) and 2 coding sequences in NADH dehydrogenase subunit 1 and subunit 5 gene in mtDNA were analyzed. MSI was only detected in 1 (2.7%), 6 (16.7%), 1 (2.9%), 1 (2.9%) or 2 (6.3%) out of 37, 36, 35, 35 or 32 cases at BAT‐25, D2S123, D3S1611, D11S904 or D17S250, respectively. LOH was found at D3S1298, D3S1561, D5S346 and TP53 in 4 (18.2%) out of 22, 2 (18.2%) out of 11, 6 (33.3%) out of 18 and 3 (12.5%) out of 24 informative cases, respectively. In mtDNA, none except a single case out of the 37 (2.7%) exhibited repeat sequence instability in the D‐loop. We conclude that the liver fluke infection‐associated ICC in Thailand is classified as low frequency MSI or microsatellite stable type and that DNA MMR system, through hMSH2 and hMLH1 gene mutations, does not play a major role in its carcinogenesis. © 2003 Wiley‐Liss, Inc.