bluuebunny/crossref_metadata_2025 huggingface dataset:
On‐line estimation of the metabolic burden resulting from the synthesis of plasmid‐encoded and heat‐shock proteins by monitoring respiratory energy generation
Authors: Hoffmann Frank , Rinas Ursula
Published: August 2002

Human basic fibroblast growth factor (hFGF‐2) was produced in high‐cell density cultures of recombinant Escherichia coli using a temperature‐inducible expression system. The synthesis rates of proteins were followed by two‐dimensional gel electrophoresis of the 35S‐methionine‐labeled proteom. After temperature induction of hFGF‐2 synthesis, the rate of total protein synthesis per biomass increased by a factor of three, mainly as a result of the additional synthesis of hFGF‐2 and heat‐shock proteins. The synthesis rates of heat‐shock proteins and constitutive plasmid‐encoded proteins increased after the temperature upshift also in the control strain without hFGF‐2 gene but followed time profiles different from the producing strain. The energy demand for the extra synthesis of plasmid‐encoded and heat‐shock proteins resulted in an elevated respiratory activity and, consequently, in a reduction of the growth rate and the biomass yield. A procedure was developed to relate the energy demand for the additional synthesis of these proteins to the generation of energy in the respiratory pathway. Specific energy production was estimated based on on‐line measurable rates of oxygen consumption, or carbondioxide evolution and growth, respectively. In this way, the metabolic burden resulting from the synthesis of plasmid‐encoded and heat‐shock proteins was quantified from on‐line accessible data. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 76: 333–340, 2001.
Direct transesterification of gases by “dry” immobilized lipase
Authors: Cameron Paula A. , Davison Brian H. , Frymier Paul D. , Barton John W.
Published: August 2002

Several different reactor configurations, including single pass, continuous recycle, and batch reactor modes, were used to investigate the effects of temperature and water activity, or relative humidity, on lipase‐catalyzed, gas‐phase transesterifications. Temperature and relative humidity were controlled both inside reactors and throughout the course of the reaction to account for and optimize their effects. Results indicated that, at low relative humidity, reaction rates increased with temperature up to 60°C. However, when relative humidity was increased, a similar increase in temperature resulted in the loss of nearly all enzyme activity. These results are consistent with the idea that enzymes without free water are more thermally stable. Furthermore, at constant ambient temperatures, production increased dramatically with an increase in relative humidity, confirming the idea that an increase in water activity increases catalytic activity. A mass balance performed on reactors at higher relative humidity revealed that hydrolysis (rather than transesterification) of the ester substrate could significantly decrease product yields. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 251–256, 2002.
Fast and efficient alkaline peroxide treatment to enhance the enzymatic digestibility of steam‐exploded softwood substrates
Authors: Yang Bin , Boussaid Abdel , Mansfield Shawn D. , Gregg David J. , Saddler John N.
Published: August 2002

The enzymatic digestibility of steam‐exploded Douglas‐fir wood chips (steam exploded at 195°C, 4.5 min, and 4.5% (w/w) SO2) was significantly improved using an optimized alkaline peroxide treatment. Best hydrolysis yields were attained when the steam‐exploded material was post‐treated with 1% hydrogen peroxide at pH 11.5 and 80°C for 45 min. This alkaline peroxide treatment was applied directly to the water‐washed, steam‐exploded material eliminating the need for independent alkali treatment with 0.4% NaOH, which has been traditionally used to post‐treat wood samples to try to remove residual lignin. Approximately 90% of the lignin in the original wood was solubilized by this novel procedure, leaving a cellulose‐rich residue that was completely hydrolyzed within 48 h, using an enzyme loading of 10 FPU/g cellulose. About 82% of the originally available polysaccharide components of the wood could be recovered. The 18% of the carbohydrate that was not recovered was lost primarily to sugar degradation during steam explosion. © 2002 John Wiley & Sons, Inc. Biotechnol Bioeng 77: 678–684, 2002; DOI 10.1002/bit.10159
A metabolic model for acetate uptake under anaerobic conditions by glycogen accumulating organisms: Stoichiometry, kinetics, and the effect of pH
Authors: Filipe Carlos D. M. , Daigger Glen T. , Grady C. P. Leslie
Published: August 2002

A metabolic model for the stoichiometry of acetate uptake under anaerobic conditions by an enriched culture of glycogen accumulating organisms (GAOs) was developed and tested by experimental studies. Glycogen served as the source of both reducing power and energy to drive the process of acetate uptake. The amount of glycogen consumed and poly‐β‐hydroxyvalerate (PHV) accumulated in the cells increased with increasing pH, indicating that the energy requirements for acetate uptake increased with pH. The composition of the accumulated poly‐β‐hydroxyalkanoates (PHAs) was adequately predicted using the assumption that acetyl‐CoA and propionyl‐CoA condense randomly to produce PHA. In addition, the rate of acetate uptake was strongly affected by the pH. The rate decreased with increasing pH and this dependence could be described with a saturation type of expression. A comparison of the rate of acetate uptake at low pH with the rates observed in enriched cultures of phosphorus accumulating organisms (PAOs) indicated that GAOs are able to compete effectively with PAOs in nutrient removal systems under certain conditions. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 76: 17–31, 2001.
Evolution of a recombinant (gucoamylase‐producing) strain of Fusarium venenatum A3/5 in chemostat culture
Authors: Wiebe Marilyn G. , Robson Geoffrey D. , Shuster Jeff , Trinci Anthony P.J.
Published: August 2002

Fusarium venenatum JeRS 325 is a transformant of strain A3/5 which produces Aspergillus niger glucoamylase (GAM) under the control of a Fusarium oxysporum trypsin‐like protease promoter. The evolution of JeRS 325 was studied in glucose‐limited chemostat cultures grown on NaNO3 or (NH4)2SO4 as the nitrogen source. Thirteen mutants which were more highly branched and four mutants which were more sparsely branched than the parental strain were isolated from the NaNO3 chemostat. The highly branched mutants detected in this chemostat did not displace the sparsely branched population. The mutants isolated from the NaNO3 chemostat complemented representative strains previously isolated from glucose‐limited chemostat cultures of F. venenatum A3/5 grown on (NH4)2SO4, but showed little complementation between themselves. By contrast, a highly branched mutant isolated from the (NH4)2SO4 chemostat culture displaced the sparsely branched mycelial population. None of the mutants isolated from the NaNO3 or (NH4)2SO4 chemostats produced as much GAM as JeRS 325. Southern blot analysis showed that all except one mutant had lost copies of both the glucoamylase and the acetamidase (the selectable marker) genes. However, specific GAM production was not necessarily correlated with the extent of glaA gene loss observed. Further, 10 of the mutants had lost the ability to grow on acetamide as the sole nitrogen source, although they retained copies of the amdS gene. In competition studies, mutants which could not utilize acetamide displaced mutants which could. The presence of foreign DNA in JeRS 325 resulted in a reduced specific growth rate (compared to A3/5), but the presence of the foreign DNA did not prevent the evolution of the strain or the isolation of mutants which had improved growth rates. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 73: 146–156, 2001.
Efficient polycyclic aromatic hydrocarbons dihydroxylation in direct micellar systems
Authors: Randazzo Demetrio , Berti Debora , Briganti Fabrizio , Baglioni Piero , Scozzafava Andrea , Di Gennaro Patrizia , Galli Enrica , Bestetti Giuseppina
Published: August 2002

Optimization of whole‐cell bioconversion of the polycyclic aromatic hydrocarbons (PAHs) anthracene, phenanthrene, and naphthalene to the enantiomerically pure corresponding cis‐dihydroxydihydro derivatives by the Escherichia coli JM109 (pPS1778) recombinant strain, carrying the naphthalene dioxygenase and corresponding regulatory genes cloned from Pseudomonas fluorescens N3, in micellar systems, is presented. We show that direct microemulsion systems, where a nonionic surfactant such as 1.5% (v/v) Triton X‐100 plus 0.6% to 1.0% (v/v) selected oils are able to solubilize the PAHs tested at relatively high concentrations (initial concentrations in the reaction medium ≥10 mM for naphthalene and phenanthrene and ≥2 mM for anthracene), and allow for more efficient substrate bioconversion. These media, while not affecting bacteria viability and performance, provide increased efficiency and final product yields (100% for naphthalene, >30% for anthracene, >60% for phenanthrene). The phase behavior of the direct microemulsion systems for the different substrates and oils utilized was monitored as a function of their volume fraction by light scattering experiments, and related to the bioconversion results. For anthracene and phenanthrene, the dihydroxylated products have an inhibitory effect on the conversion reactions, thus hindering complete turnover of the substrates. We ascertain that such inhibition is reversible because removal of the products formed allowed the process to start over at rates comparable to initial rates. To allow for complete conversion of the PAHs tested a stepwise or continuous separation of the product formed from the micellar reaction environment is being developed. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 240–248, 2001.
Capillary isoelectric focusing–mass spectrometry: analysis of protein mixtures from human body fluids
Authors: Clarke Nigel J. , Naylor Stephen
Published: August 2002

Isoelectric focusing within a fused silica capillary (cIEF) has proved to be a powerful and practical method for high‐resolution separation of analytes from complex biological mixtures. This technique overcomes many of the problems of isoelectric focusing within slab gel media. However current cIEF systems commonly utilize UV detection which limits the detail of analyte structural information that is obtained during analysis. The use of mass spectrometry (MS) as the detection system provides much greater structural information about the detected analytes allowing accurate relative molecular mass (Mr) determination for proteins and polypeptides. We have constructed a cIEF‐MS interface and compared the separation of standard proteins analyzed by cIEF‐UV with cIEF‐MS. This allowed rapid optimization of the cIEF‐MS system performance. Further we have demonstrated the use of MS as a detection system provides accurate Mr information and can provide analyte modification details. These factors increase the likelihood of absolute identification for physiological proteins within complex in vivo‐derived mixtures. To demonstrate the value of cIEF‐MS in such analyses we have undertaken an examination of cerebrospinal fluid (CSF), and tentatively identified a number of constituent proteins. We have also analyzed whole blood from control and diabetic patients. We show that glycated α‐ and β‐ chains of hemoglobin are found in almost equal abundance in diabetic patient blood. From these results we suggest cIEF‐MS is an efficient and useful tool for the separation and examination of in vivo‐derived analytes within physiological fluids. Copyright © 2002 John Wiley & Sons, Ltd.
A fluorimetric, column‐switching HPLC and its application to an elimination study of LLU‐α enantiomers in rat plasma
Authors: Hattori Akihiro , Fukushima Takeshi , Hamamura Kimio , Kato Masaru , Imai Kazuhiro
Published: August 2002

A method for the enantiomeric determination of 2,7,8‐trimethyl‐2‐(β‐carboxyethyl)‐6‐hydroxy chroman (LLU‐α, γ‐CEHC) in rat plasma was developed using high‐performance liquid chromatography (HPLC) with a fluorimetric derivatization with 4‐N, N‐dimethylaminosulfonyl‐7‐piperazino‐2,1,3‐benzoxadiazole (DBD‐PZ) followed by O‐acetylation with acetyl chloride. The proposed HPLC system used two non‐chiral columns (phenyl and octadecylsilica) and a chiral column (a modified cellulose type), which were connected via two column‐switching valves. A derivatized sample prepared from rat plasma was first separated on the phenyl column, and the fraction including LLU‐α derivative was introduced to the octadecylsilica column to quantify the concentration of the mixture of S‐ and R‐LLU‐α. Finally, the LLU‐α derivative was directly injected into the chiral column to obtain the ratio of the enantiomers. The proposed HPLC system was applied to the enantiomeric determination of LLU‐α in plasma after intravenous administration of racemic LLU‐α. S‐LLU‐α was eliminated faster than R‐LLU‐α, and its concentration in plasma decreased to one‐third at 2 min after dosing. Copyright © 2001 John Wiley & Sons, Ltd.

Abbreviations used:

DBD‐PZ

4‐N,N‐dimethylaminosulfonyl‐7‐piperazino‐2,1,3‐benzoxadiazole

DPDS

2,2′‐dipyridinyl disulfide

LLU‐α, γ‐CEHC

2,7,8‐trimethyl‐2‐(β‐carboxyethyl)‐6‐hydroxy chroman

TFA

trifluoroacetic acid

TPP

triphenylphosphine
Knowledge interests in corporate environmental management
Authors: Dobers Peter , Strannegård Lars , Wolff Rolf
Published: August 2002

We know from earlier studies that corporate environmental management is a young discipline, not yet integrated in general management and organization studies, but that researchers take an increasing part in the ongoing scientific conversation. However, the underlying knowledge interests characterizing the field of corporate environmental management is yet empirically unsubstantiated. One way to find out what elements make up the field is to analyse the contents of the most influential writings in the field. The present article identifies the 10 most cited works in Business Strategy and the Environment in 1992–2000 and explores the content of these texts. We conceptualise a typology for analysing corporate environmental management theory and formulate a characterization of the dominating knowledge interests. Our findings show that the theoretical fundament of corporate environmental management lacks a hermeneutic knowledge interest. Copyright © 2001 John Wiley & Sons, Ltd and ERP Environment
The evolution of environmental management: from stage models to performance evaluation
Authors: Kolk Ans , Mauser Anniek
Published: August 2002

In the past two decades, academics and practitioners have attempted to improve understanding of environmental management by classifying companies' environmental behaviour, and evaluating their performance. Driven by both research and societal interest, this has resulted in a wave of stage or phase models, and a range of typologies. This article gives an overview of the development of such environmental management models, analysing their characteristics, strengths and weaknesses. An evolution can be noted in the direction of typologies and non‐linear models to deal with organizational and strategic complexities. Models are starting to pay more attention to the management side. To overcome problems of operationalization and limited company and sector specificity, environmental performance evaluation systems have emerged more recently. Although comprehensive performance assessments are still unavailable, the tenets of such a system can already be delineated. The paper presents these components, and draws conclusions on the contribution of environmental management models and performance evaluation systems. Copyright © 2002 John Wiley & Sons, Ltd. and ERP Environment
Pretarsal fat compartment in the lower eyelid
Authors: Hwang Kun , Joong Kim Dae , Chung Robert S.
Published: August 2002

It is generally accepted that there are three infraorbital fat regions in the lower eyelid; medial, central, and lateral compartments. However, removing only the fat in the lateral compartment does not remove the bulge just below the eyelashes, which is caused by another fat pad. The aim of this study was to describe the anatomy of the pretarsal fat compartment and to demonstrate its clinical implications in lower lid blepharoplasty. Ten cadavers (total 20 lower eyelids) were studied. A skin‐muscle flap was reflected to expose the soft pretarsal structures. A small stab incision was made on the lateral portion of the sac containing fat on the tarsus. Methylene blue dye was injected into the sac. Specimens were fixed and sagittal sections in four different planes were prepared for histological analysis. The injected dye remained within the sac and demarcated it as a pear or cone shaped structure. This encapsulated fat compartment sits on the lateral half of the tarsal plate above the lateral compartment fat. Auxillary or submuscular fat is well known. This study, however, designates the pretarsal fat as “encapsulated” in a compartment instead of being unbound. We have named it the “pretarsal fat compartment.” Histologically, orbital septal fibers separate “pretarsal fat” from lateral infraorbital fat. It is recommended that fat in the pretarsal fat compartment be removed during lower lid blepharoplasty in order to alleviate the bulge or knoll of the skin just below the lower eyelashes. Clin. Anat. 14:179–183, 2001. © 2001 Wiley‐Liss, Inc.
Eicosapentaenoic acid and docosahexaenoic acid effects on tumour mitochondrial metabolism, acyl CoA metabolism and cell proliferation
Authors: Colquhoun Alison , Ramos Karina L. , Schumacher Robert I.
Published: August 2002

In order to investigate the effects of high‐fat diets rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), Wistar rats bearing subcutaneous implants of the Walker 256 tumour were fed pelleted chow containing low DHA/EPA or high DHA/EPA. The presence of n‐3 polyunsaturated fatty acids (PUFAs) led to a marked suppression (35–46%) of tumour growth over a 12 day period. Both the whole tumour homogenate and the Percoll‐purified mitochondrial fraction presented significant changes in fatty acid composition. The levels of EPA increased in both n‐3 dietary groups while the levels of DHA increased only in the high DHA/EPA group, in comparison with the control chow‐fed group. The presence of n‐3 PUFAs led to an increase in mitochondrial acyl CoA synthetase activity, but neither the cytoplasmic acyl CoA content nor the n‐3 fatty acid composition of the cytoplasmic acyl CoAs was altered by the diet. The content of thiobarbituric acid‐reactive substances (TBARS) was increased in the low DHA/EPA group but was unchanged in the high DHA/EPA group. In vitro studies with the Walker 256 cell line showed a 46% decrease in cell growth in the presence of either EPA or DHA which was accompanied by a large decrease in the measured mitochondrial membrane potential. The TBARS content was increased only in the EPA‐exposed cells. Cell cycle analysis identified a decrease in G0–G1 phase cells and an increase in G2–M phase cells and apoptotic cells, for both EPA and DHA‐exposed cells. The data show that the presence of n‐3 PUFAs in the diet is able to significantly after the growth rate of the Walker 256 tumour. The involvement of changes in mitochondrial membrane composition and membrane potential have been indicated for both EPA and DHA, while changes in lipid peroxidation have been identified in the presence of EPA but not of DHA. Copyright © 2001 John Wiley & Sons, Ltd.
The uncoupling effect of the nonsteroidal anti‐inflammatory drug nimesulide in liver mitochondria from adjuvant‐induced arthritic rats
Authors: Caparroz‐Assef S. M. , Salgueiro‐Pagadigorria C. L. , Bersani‐Amado C. A. , Bracht A. , Kelmer‐Bracht A. M. , Ishii‐Iwamoto E. L.
Published: August 2002

The aim of the present study was to evaluate the changes caused by adjuvant‐induced arthritis in liver mitochondria and to investigate the effects of the nonsteroidal anti‐inflammatory drug nimesulide. The main alterations observed in liver mitochondria from arthritic rats were: higher rates of state IV and state III respiration with β‐hydroxybutyrate as substrate; reduced respiratory control ratio and impaired capacity for swelling dependent on β‐hydroxybutyrate oxidation. No alterations were found in the activities of NADH oxidase and ATPase. Nimesulide produced: (1) stimulation of state IV respiration; (2) decrease in the ADP/O ratio and in the respiratory control ratio; (3) stimulation of ATPase activity of intact mitochondria; (4) inhibition of swelling driven by the oxidation of β‐hydroxybutyrate; (5) induction of passive swelling due to NH3/NH4+ redistribution. The activity of NADH oxidase was insensitive to nimesulide. Mitochondria from arthritic rats showed higher sensitivity to nimesulide regarding respiratory activity. The results of this work allow us to conclude that adjuvant‐induced arthritis leads to quantitative changes in some mitochondrial functions and in the sensitivity to nimesulide. Direct evidence that nimesulide acts as an uncoupler was also presented. Since nimesulide was active in liver mitochondria at therapeutic levels, the impairment of energy metabolism could lead to disturbances in the liver responses to inflammation, a fact that should be considered in therapeutic intervention. Copyright © 2001 John Wiley & Sons, Ltd.
Literature on Community Colleges: An Overview
Authors: Bers Trudy H. , Calhoun Harriott D.
Published: August 2002

The literature on community colleges is varied in terms of categories, types, purposes, authors, and audiences. It reveals a gap between the perceptions of the research community and perceptions among practitioners regarding the important topics for research and publication.
Recanalization and thrombectomy of internal anastomosis in a patient with tetralogy of Fallot using the AngioJet rheolytic catheter
Authors: Kokov L.S. , Korostelev A.N. , Grinko A.N. , Ilyina M.V. , Solopova G.V. , Aiazian E. , Setum C.M.
Published: August 2002

The modified Blalock‐Taussig shunt is the palliative treatment of choice for tetralogy of Fallot. Shunt thrombosis is a potential complication, requiring high‐risk reoperation. The use of percutaneous rheolytic devices for thrombus removal in such occluded shunts has not been previously reported. We describe a case in which use of a rheolytic catheter resulted in significant thrombus removal and rapid reversal of cyanosis and dyspnea in a 5‐year‐old patient. The patient remains free of symptoms at 30‐day follow‐up. Cathet Cardiovasc Intervent 2001;53:504–507. © 2001 Wiley‐Liss, Inc.
Intra‐aortic counterpulsation during carotid stenting
Authors: Hennen Benno , Gröschel‐Guth Angelika , Scheller Bruno , Hammer Bernd , Böhm Michael
Published: August 2002

Postprocedural hypotension following endovascular stent placement of carotid artery disease (CAS) predicts increased in‐hospital complications and long‐term risk of death. Intra‐aortic balloon counterpulsation (IABP) both increases mean arterial pressure and cerebral blood flow and therefore possibly reduces complications due to hemodynamic instability during and after CAS. In this study, we describe the use of IABP in a patient with severe depression of left ventricular function due to diffuse coronary artery disease undergoing CAS. Controlled studies are necessary to demonstrate a potentially protective role of IABP in high‐risk CAS patients. Cathet Cardiovasc Intervent 2001;53:546–548. © 2001 Wiley‐Liss, Inc.
The patentability of biomolecules – does online bioinformatics compromise novelty?
Authors: Hutter Anton
Published: August 2002

Researchers are becoming increasingly concerned that the confidentiality
of their novel biomolecule sequences is being jeopardised, particularly when
these sequences are either submitted to sequence databases or uploaded as
query terms onto internet‐based bioinformatic software suites. The researcher's
fears stem from the fact that the actual uploading of their sequences acts
as a novelty destroying prior disclosure or publication, and that this may
subsequently preclude valid patent protection for the sequences. This article
addresses the key issues involved in the analyses of biomolecules, highlighting
potential risks taken by many researchers in regard to patent protection and
suggests possible ways in which these risks may be mitigated. Copyright ©
2002 John Wiley & Sons, Ltd.
Featured Organism: Arabidopsis thaliana
Authors: Wixon Jo
Published: August 2002

Arabidopsis is universally acknowledged as the model for dicotyledonous crop plants. Furthermore, some of the information gleaned from this small plant can be used to aid work on monocotyledonous crops. Here we provide an overview of the current state of knowledge and resources for the study of this important model plant, with comments on future prospects in the field from Professor Pamela Green and Dr Sean May. Copyright © 2001 John Wiley & Sons, Ltd.

Summary

•  Small flowering plant of mustard family (Brassicaceae)

•  Rapid life cycle, high seed production, easy to cultivate

•  Small genome, 125 Mb, as five chromosomes

•  Genome sequence completed, 25,498 genes identified

•  30% of genes currently have no functional classification

•  Majority of genome present in two copies, suggesting whole genome duplication
Optical resolution of 6‐fluoro‐2‐methyl‐1,2,3,4‐tetrahydroquinoline by supercritical fluid extraction
Authors: Kmecz Ildikó , Simándi Béla , Bálint József , Székely Edit , Fogassy Elemér , Kemény Sándor
Published: August 2002

6‐Fluoro‐2‐methyl‐1,2,3,4‐tetrahydroquinoline (FTHQ) enantiomers were separated by supercritical fluid extraction using carbon dioxide. Diastereoisomeric salts were formed from the racemic base with less than one equivalent of O,O′‐di‐(4‐toluoyl)‐(2R,3R)‐tartaric acid (DPTTA). Further purification was achieved by partial salt formation of the enantiomeric mixture with an achiral acid (HCl) followed by the supercritical fluid extraction of the free enantiomers. Chirality 13:568–570, 2001. © 2001 Wiley‐Liss, Inc.
Enantiomeric recognition of chiral 3,3‐bridged‐1,1′‐binaphthol dimer toward α‐phenylethylamine and α‐amino acid ester
Authors: Liu Tian Jun , Chen Yong Jun , Zhang Ke Sheng , Wang Dong , Guo Da Wei , Yang Xiao Zhen
Published: August 2002

The 1,1′‐binaphthol‐based dimers with p‐phenylenebis(2‐ethynyl) spacer, (+)‐6 and (+)‐2, were synthesized as chiral host compounds. 1H NMR, UV‐vis, and fluorescent titration were used to evaluate the enantiomeric recognition abilities of the chiral host dimers toward the guest amine 7 and α‐amino acid ester 8. The chiral BINOL‐based dimers were found to have good enantiomeric recognition ability. The computer simulation of the host–guest complex molecules was carried out to describe the conformational changes of both naphthyl ring in the molecule of chiral host dimer after complexation with the guest molecule. Chirality 13:595–600, 2001. © 2001 Wiley‐Liss, Inc.
Reorganization of the actin cytoskeleton in the protruding lamellae of human fibroblasts
Authors: Safiejko‐Mroczka Barbara , Bell Paul B.
Published: August 2002

To investigate the mechanisms of protrusion in vertebrate cells, the primary event in cell motility, human fibroblasts were treated with neomycin, an inhibitor of the phosphatidylinositol cycle, to induce protrusion. Changes in cell motility and the cytoskeleton were examined by video, fluorescence, scanning electron, and confocal microscopy and by cytofluorometry. Protrusion in neomycin‐treated human fibroblasts is correlated with a transient overall decrease in F‐actin followed by an increase in F‐actin at the leading edge of the protruding lamella. In growing lamellae, F‐actin is organized in a marginal band at the leading edge. Although actin is present in the lamella behind the leading edge, very little of it is F‐actin. Scanning electron microscopy of detergent‐extracted cells reveals a band of dense filaments at the leading edge, corresponding to the marginal band of F‐actin seen in fluorescently labeled cells, and a sparse population of short, fragmented filaments, in the rest of the lamella. Gelsolin is colocalized with F‐actin in the marginal band and is also present in the lamella where F‐actin is largely absent. The data support the hypothesis that the protrusion is initiated by the breakdown of cortical actin filaments, possibly mediated by gelsolin, whereas expansion of the protrusion requires de novo polymerization of actin filaments at the leading edge. Cell Motil. Cytoskeleton 50:13–32, 2001. © 2001 Wiley‐Liss, Inc.
Magnetic resonance imaging compatible neonate incubator
Authors: Dumoulin Charles L. , Rohling Kenneth W. , Piel Joseph E. , Rossi Charles J. , Giaquinto Randy O. , Watkins Ronald D. , Vigneron Daniel B. , Barkovich A. James , Newton Nancy
Published: August 2002

An incubator for magnetic resonance imaging (MRI) of premature and full‐term infants was designed and constructed. The device is an extensively modified MRI system table that has a permanently attached incubator housing, self‐contained power supply, and full patient monitoring capability. A custom eight‐element birdcage coil within the incubator slides over the infant and provides high sensitivity for MRI. The incubator provides all the temperature control and patient monitoring found in conventional neonatal intensive care incubators, but in an MRI‐compatible fashion. The table can operate for at least three hours on a single battery charge. It is believed that this table design will enable routine MRI scanning of newborn infants requiring extensive monitoring of vital signs and/or temperature regulation. © 2002 Wiley Periodicals, Inc. Concepts in Magnetic Resonance (Magn Reson Engineering) 15: 117–128, 2002
HER‐2/neu overexpression is rare in hepatocellular carcinoma and not predictive of anti‐HER‐2/neu regulation of cell growth and chemosensitivity
Authors: Hsu Chiun , Huang Chin‐Lun , Hsu Hey‐Chi , Lee Po‐Huang , Wang Shiou‐Jeng , Cheng Ann‐Lii
Published: August 2002

BACKGROUND

The overexpression of HER‐2/neu oncogene has been implicated in the development and modulation of many types of cancer. However, whether HER‐2/neu overexpression plays a similar role in hepatocellular carcinoma (HCC) has not been determined.

METHODS

Tissue specimens from 36 HCC patients who had been enrolled in 3 separate prospective clinical trials of systemic chemotherapy were studied by immunohistochemical staining. A polyclonal antibody (A0485; DAKO, Copenhagen, Denmark) against HER‐2/neu and a horseradish peroxidase‐based visualization system (Envision+, DAKO) was used. Scoring criteria was in accordance with the manufacturer's guidelines. Twelve HCC cell lines were examined for HER‐2/neu overexpression by Western blotting. Single‐agent growth regulatory activity of the anti–HER‐2/neu antibody, trastuzumab (Herceptin; Genentech, South San Francisco, CA), and its combinative cytotoxicity with chemotherapeutic agents (doxorubicin, gemcitabine, cisplatin, irinotecan) were determined by a tetrazolium‐based colorimetric assay (MTT test).

RESULTS

All but one of the HCC tumor tissues were negative for HER‐2/neu expression. The only patient with positive HER‐2/neu expression was a 57‐year‐old male patient who achieved stabilization of disease for 2 months after chemotherapy. Eight of the 35 patients with negative HER‐2/neu expression had had partial remission after chemotherapy (P = 0.78). Only one (Tong cells) of the 12 HCC cell lines had a significant level of HER‐2/neu expression. However, trastuzumab up to 10 μg/mL had no discernible growth inhibitory or chemosensitizing effect on Tong cells or any other cell lines.

CONCLUSIONS

HER‐2/neu overexpression is rare in human HCC tissues, and anti–HER‐2/neu regulation appears to play little role in the treatment of this tumor. Cancer 2002;94:415–20. © 2002 American Cancer Society.
European Organization for Research and Treatment of Cancer and Groupe d'Etude des Lymphomes de l'Adulte very favorable and favorable, lymphocyte‐predominant Hodgkin disease
Authors: Wilder Richard B. , Schlembach Pamela J. , Jones Dan , Chronowski Gregory M. , Ha Chul S. , Younes Anas , Hagemeister Fredrick B. , Barista Ibrahim , Cabanillas Fernando , Cox James D.
Published: August 2002

BACKGROUND

Lymphocyte‐predominant Hodgkin disease (LPHD) is rare and has a natural history different from that of classic Hodgkin disease. There is little information in the literature regarding the role of chemotherapy in patients with early‐stage LPHD. The objective of this study was to examine recurrence free survival (RFS), overall survival (OS), and patterns of first recurrence in patients with LPHD who were treated with radiotherapy alone or with chemotherapy followed by radiotherapy.

METHODS

From 1963 to 1996, 48 consecutive patients ages 16–49 years (median, 28 years) with Ann Arbor Stage I (n = 30 patients) or Stage II (n = 18 patients), very favorable (VF; n = 5 patients) or favorable (F; n = 43 patients) LPHD, according to the European Organization for Research and Treatment of Cancer and Groupe d'Etude des Lymphomes de l'Adulte (EORTC‐GELA) criteria, received radiotherapy alone (n = 37 patients) or received chemotherapy followed by radiotherapy (n = 11 patients). The percentages of patients with VF disease (11% vs. 9% in the radiotherapy group vs. the chemotherapy plus radiotherapy group, respectively) or F disease (89% vs. 91%, respectively) within the two treatment groups were similar (P = 1.00). A median of three cycles of chemotherapy with mechlorethamine, vincristine, procarbazine, and prednisone (MOPP) or with mitoxantrone, vincristine, vinblastine, and prednisone (NOVP) was given initially to six patients and five patients, respectively. A median total radiotherapy dose of 40 grays (Gy) given in daily fractions of 2.0 Gy was delivered to both treatment groups.

RESULTS

The median follow‐up was 9.3 years, and 98% of patients were observed for ≥ 3.0 years. RFS was similar for patients who were treated with radiotherapy alone and patients who were treated with chemotherapy followed by radiotherapy (10‐year survival rates: 77% and 68%, respectively; P = 0.89). The OS rate also was similar for the two groups (10‐year survival rates: 90% and 100%, respectively; P = 0.43). MOPP or NOVP chemotherapy did not reduce the risk of recurrence outside of the radiotherapy fields.

CONCLUSIONS

MOPP or NOVP chemotherapy did not improve RFS or OS significantly in patients with VF or F LPHD, although the statistical power was limited. Ongoing clinical trials will help to clarify the role of a watch‐and‐wait strategy or systemic therapy, including anthracycline (epirubicin or doxorubicin), bleomycin, and vinblastine‐based chemotherapy or antibody‐based approaches, in the treatment of these patients. Cancer 2002;94:1731–8. © 2002 American Cancer Society.

DOI 10.1002/cncr.10404
Expression of estrogen receptor, progesterone receptor, and insulin‐like growth factor receptor‐1 and of MIB‐1 in patients with recurrent pleomorphic adenoma of the parotid gland
Authors: Glas Afina S. , Hollema Harry , Nap Raoul E. , Plukker John T.
Published: August 2002

BACKGROUND

Patients with recurrent pleomorphic adenomas of the parotid gland are difficult to manage without considerable risk of facial nerve injury. The prognostic significance of progesterone receptor (PR) and estrogen receptor (ER) reported in these adenomas was evaluated in patients with recurrent pleomorphic adenomas, comparing the results in a group of patients with primary adenomas without recurrences during 10 years of follow‐up.

METHODS

Paraffin embedded tumor samples from 52 patients with recurrent pleomorphic adenoma of the parotid gland were collected and stained immunohistochemically. Expression of PR, ER, Ki‐67 antigen, and insulin‐like growth factor receptor‐1 (IGFR‐1) was analyzed in resected samples of recurrent tumors and was compared with samples from a control group of patients with primary pleomorphic adenoma.

RESULTS

A difference (P < 0.05) in the type of tumor was observed between the recurrent group (more cell‐poor variants) and the control group. ER expression was low in both groups (19% and 17%, respectively), but immunoreactivity for ER was higher (48%) in normal parotid gland tissue. PR expression in the recurrent group (96%) was higher compared with PR expression in the control group (61%; P < 0.001). PR expression and IGFR‐1 expression were correlated weakly (correlation coefficient = 0.660; P = 0.053) in the recurrent group. The expression of growth fraction (Ki‐67 score) and IGFR‐1 was similar in both groups but was more extensive compared with normal parotid gland tissue.

CONCLUSIONS

PR seems to be a prognostic factor in recurrent pleomorphic adenoma of the parotid gland. The PR pathway can be considered a potential target for hormone treatment in patients with these recurrent adenomas. Cancer 2002;94:2211–16. © 2002 American Cancer Society.

DOI 10.1002/cncr.10445
Basic organization of projections from the oval and fusiform nuclei of the bed nuclei of the stria terminalis in adult rat brain
Authors: Dong Hong‐Wei , Petrovich Gorica D. , Watts Alan G. , Swanson Larry W.
Published: August 2002

The organization of axonal projections from the oval and fusiform nuclei of the bed nuclei of the stria terminalis (BST) was characterized with the Phaseolus vulgaris‐leucoagglutinin (PHAL) anterograde tracing method in adult male rats. Within the BST, the oval nucleus (BSTov) projects very densely to the fusiform nucleus (BSTfu) and also innervates the caudal anterolateral area, anterodorsal area, rhomboid nucleus, and subcommissural zone. Outside the BST, its heaviest inputs are to the caudal substantia innominata and adjacent central amygdalar nucleus, retrorubral area, and lateral parabrachial nucleus. It generates moderate inputs to the caudal nucleus accumbens, parasubthalamic nucleus, and medial and ventrolateral divisions of the periaqueductal gray, and it sends a light input to the anterior parvicellular part of the hypothalamic paraventricular nucleus and nucleus of the solitary tract. The BSTfu displays a much more complex projection pattern. Within the BST, it densely innervates the anterodorsal area, dorsomedial nucleus, and caudal anterolateral area, and it moderately innervates the BSTov, subcommissural zone, and rhomboid nucleus. Outside the BST, the BSTfu provides dense inputs to the nucleus accumbens, caudal substantia innominata and central amygdalar nucleus, thalamic paraventricular nucleus, hypothalamic paraventricular and periventricular nuclei, hypothalamic dorsomedial nucleus, perifornical lateral hypothalamic area, and lateral tegmental nucleus. Moderately dense inputs are found in the parastrial, tuberal, dorsal raphé, and parabrachial nuclei and in the retrorubral area, ventrolateral division of the periaqueductal gray, and pontine central gray. Light projections end in the olfactory tubercle, lateral septal nucleus, posterior basolateral amygdalar nucleus, supramammillary nucleus, and nucleus of the solitary tract. These and other results suggest that the BSTov and BSTfu are basal telencephalic parts of a circuit that coordinates autonomic, neuroendocrine, and ingestive behavioral responses during stress. J. Comp. Neurol. 436:430–455, 2001. © 2001 Wiley‐Liss, Inc.
Expression of active caspase‐3 in mitotic and postmitotic cells of the rat forebrain
Authors: Yan Xiao‐Xin , Najbauer Joseph , Woo Cynthia C. , Dashtipour Khashayar , Ribak Charles E. , Leon Michael
Published: August 2002

Active caspase‐3 immunoreactivity was detected in the rat forebrain proliferative regions at birth and remained high in these areas for about 2 weeks, during which period labeled cells were present centroperipherally across the olfactory bulb. By the end of the third postnatal week, only a small number of immunolabeled cells remained in these forebrain structures. Active caspase‐3 immunolabeling was localized mostly to cell nuclei and co‐localized partially with TuJ1 and NeuN immunoreactivity, but not with glial fibrially acidic protein, OX‐42, γ‐aminobutyric acid, or terminal deoxynucleotidyl transferase‐mediated nick end labeling (TUNEL)‐positive labeling. Active caspase‐3 and 5‐bromo‐2′‐deoxyuridine (BrdU) double‐labeled nuclei were seen in the proliferative regions after 2 hours and in the periglomerular region of the bulb after 7 days following BrdU injections. Examination of the cells with electron microscopy confirmed that the active caspase‐3‐containing nuclei in the proliferative regions often had infoldings and appeared to be undergoing division. Some of the cells with active caspase‐3‐labeled nuclei in the bulb had synapses on their somata or dendrites. Labeled dendritic spines and a few axon terminals were also observed in the olfactory bulb. Taken together, it appears that a wave of active caspase‐3‐positive cells are dividing in the proliferative zones and then migrating to the bulb as they differentiate into neurons. Therefore, active caspase‐3 may play a role in cellular processes such as neuronal differentiation, migration, and plasticity, in addition to its role in cell death. J. Comp. Neurol. 433:4–22, 2001. © 2001 Wiley‐Liss, Inc.
3‐D mixed‐mode K‐calculations with the interaction integral method and the quarter point element stress method
Authors: Dhondt G.
Published: August 2002

The K‐distribution along a straight crack in a single edge notch specimen and a slant crack in a three‐point bending specimen is determined using the interaction integral method (IINT) and the quarter point element stress method (QPES). The results are discussed and recommendations for the proper use of both methods are given. Copyright © 2001 John Wiley & Sons, Ltd.
A new triangular element for the analysis of thick and thin plates
Authors: Sheikh A. H. , Dey P.
Published: August 2002

A new triangular element based on Reissner–Mindlin's plate theory is presented. It has three corner nodes and three mid‐side nodes where each node contains three usual degrees of freedom, i.e. transverse displacement (w) and rotations of the normal to the plate middle plane due to bending and shear (θx and θy). The element formulation is based on displacement approach and it is done in such a manner, which has made the element free from shear locking and spurious modes. This is achieved through proper selection of displacement field and its representation. It contains transverse displacement (w) and rotations of normal due to shear (γx and γy). The concept of taking γx and γy as independent displacement components instead of θx and θy has helped to develop such an element. The performance of the element is tested through numerical examples. Copyright © 2001 John Wiley & Sons, Ltd.
Accounting for stability: a posteriori error estimates based on residuals and variational analysis
Authors: Estep Donald , Holst Michael , Mikulencak Duane
Published: August 2002

A posteriori error estimates have had a major impact on adaptive error control for the finite element method. In this paper, we review a relatively new approach to a posteriori error estimation based on residuals and a variational analysis. This approach is distinguished by a direct attempt to account for the effects of stability on the propagation of error. We illustrate properties of this approach using several examples. Copyright © 2001 John Wiley & Sons, Ltd.
Spectral power distributions for the CIE stimuli
Authors: Hawkyard C. J. , de M Bezerra C.
Published: August 2002

The CIE reference colour stimuli, X, Y, and Z, were derived by constructing a triangle outside the R,G,B triangle and outside the area bounded by the spectrum locus and the purple line. By this means, all colours, including monochromatic ones, have positive tristimulus values. The colour‐matching functions are the relative quantities of these stimuli required to be mixed additively to match the equal energy monochromatic colours. The stimuli are not realizable as light sources, and the CIE has not specified their spectral power distributions. There is an infinite number of spectral power distributions whose properties meet the prerequisites for X (X = 100, Y = 0, Z = 0), Y (0, 100, 0), and Z (0, 0, 100), and two possible sets have been calculated by different methods. These curves could be used as primary red, green, and blue lights in additive mixing to produce synthetic reflectance curves, which are useful in the specification of on‐screen colours, and as a means of producing colour constant standards. © 2001 John Wiley & Sons, Inc. Col Res Appl, 26, 478–482, 2001
Strong types for coordinating active objects
Authors: Puntigam Franz
Published: August 2002

An object type is usually regarded as a contract between an object and each of its clients. However, in concurrent (and sometimes also in sequential) systems it is more useful to regard a type as a contract between an object and the set of all clients: when the object acts as a shared resource, the clients must be coordinated before sending messages to the object. Process types of active objects specify constraints on the ordering of messages. Static type checking ensures proper coordination of clients so that objects receive messages only in acceptable orderings. As presented in this article, the process type system is based on a simple computation model where active objects communicate via asynchronous message passing. Copyright © 2001 John Wiley & Sons, Ltd.
Determination of individual cell Michaelis‐Menten constants
Authors: Sunray Merav , Zurgil Naomi , Shafran Yana , Deutsch Mordechai
Published: August 2002

Background

A novel methodology for the measurement and analysis of apparent KM (Michaelis‐Menten constant) and VMAX values of individual cells is suggested. It is based on a mathematical model that considers substrate influx into the cell, its intracellular enzymatic hydrolysis, and the product efflux. The mathematical formulation was approximated linearly in order to analyze intracellular substrate conversion characteristics via Michaelis‐Menten theory.

Methods

Utilizing static cytometry, the time dependence of the fluorescence intensity [FI(t)] emitted from prelocalized and defined FDA stained cells was recorded. This required frequent periodical measurements of the same cells, which are sequentially exposed to various fluorogenic substrate concentrations.

Results

Model simulations correlated with experimental results. Differences in distributions of individual KM and VMAX values of cells incubated with and without PHA were evident. Average KM and VMAX values of PHA‐stimulated cells increased by 99% and 540%, respectively.

Conclusions

This study may provide a tool for assessing intracellular enzymatic activity in individual intact cells under defined physiologic conditions. This may open new vistas in various areas, giving answers to critical questions arising in the field of cell and developmental biology, immunology, oncology, and pharmacology. Cytometry 47:8–16, 2002. © 2001 Wiley‐Liss, Inc.
Crush preparation findings of “sarcomatoid” chordoma of the sacrum: Report of a case with histologic, immunohistochemical, and ultrastructural correlation
Authors: Ng W.K. , Tang V.
Published: August 2002

We report on the crush preparation findings of a case of “sarcomatoid” chordoma occurring in the sacral region of a 78‐yr‐old Chinese male. The smears showed clumps and small cords of polygonal tumor cells containing bubbly cytoplasm and round to oval nuclei. Focally, there were also aggregates of long filamentous spindle cells and stellate bizarre cells with marked nuclear pleomorphism. Occasional tumor cells were seen in association with dense amorphous material. Histologic examination of the excised specimen showed features of the so‐called “sarcomatoid” chordoma which consisted of prominent foci of mitotically inactive spindle and pleomorphic cells, in addition to the conventional chordoma areas. An osteosarcoma‐like pattern of probably metaplastic nature was also seen within the tumor. Immunohistochemical study showed that most tumor cells expressed cytokeratins. Ultrastructural examination revealed the characteristic rough endoplasmic reticulum‐mitochondria complexes. While there are many spindle and pleomorphic cells seen in crush preparations, the distinction from other true high‐grade malignancies is important. Recognition of these “pseudoanaplastic” cytologic features also helps to expand the morphologic spectrum of chordoma. Diagn. Cytopathol. 25:406–410, 2001. © 2001 Wiley‐Liss, Inc.