bluuebunny/crossref_metadata_2025 huggingface dataset:
Activity of different Candida antarctica lipase B formulations in organic solvents
Authors: Secundo Francesco , Carrea Giacomo , Soregaroli Carla , Varinelli Donatella , Morrone Raffaele
Published: August 2002

The activity of different formulations of Candida antarctica lipase B (CALB), such as crude CALB, purified CALB, purified CALB lyophilized with PEG (CALB + PEG) or oleic acid (CALB + OA), and the commercial formulation Novozym 435, was determined in toluene, carbon tetrachloride, and 1,4‐dioxane at various water activities (aw). The reaction between vinylacetate and 1‐octanol was used as the model reaction and both transesterification (formation of 1‐octylacetate) and hydrolytic (formation of acetic acid from vinylacetate) activities were determined. For equal amounts of lipase protein, CALB + PEG (and to a lesser extent CALB + OA) displayed higher activity than that of the other formulations; for instance, in toluene (aw < 0.1), it was 260‐, 13‐, and 1.8‐fold more active than crude CALB, purified CALB, and Novozym 435, respectively. Moreover, the transesterification activity of CALB + PEG was of the same order of magnitude (51%) of the activity shown by the enzyme in the hydrolysis of vinylacetate in aqueous buffer. These results suggest that PEG and oleic acid could act as lyoprotectants, preventing the formation of intermolecular interactions during the lyophilization process that might be responsible for protein denaturation. No diffusional limitation was observed for CALB + PEG‐catalyzed reactions. Purified CALB, in contrast to the other formulations, showed a marked activity increase (2.1 to 7.8‐fold) as a function of aw and, in 1,4‐dioxane, it was 3.5‐fold more active when it was added to the solvent after previous dissolution of the lyophilized powder in water. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 73: 157–163, 2001.
Highly efficient synthesis of ampicillin in an “aqueous solution‐precipitate” system: Repetitive addition of substrates in a semicontinuous process
Authors: Youshko Maxim I. , van Langen Luuk M. , de Vroom Erik , van Rantwijk Fred , Sheldon Roger A. , Švedas Vytas K.
Published: August 2002

The synthesis of ampicillin catalyzed by Escherichia coli penicillin acylase was optimized in an aqueous system with partially dissolved antibiotic nucleus 6‐aminopenicillanic acid (6‐APA). The yields of both 6‐APA and acyl donor could be improved by repetitively adding substrates to the reaction, allowing the concentration of 6‐APA to remain saturated throughout. In this reaction concept, with four subsequent additions of substrates, 97% conversion of 6‐APA and 72% of D‐(−)‐phenylglycine methyl ester (D‐PGM) to ampicillin was achieved. The synthetic potential of this concept was estimated using a mathematical model which showed that by increasing the amount of added substrates a nearly quantitative conversion of 6‐APA and 85% conversion of acyl donor into ampicillin could be achieved . © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 73: 426–430, 2001.
Limited adhesion of biodegradable microspheres to E‐ and P‐selectin under flow
Authors: Dickerson J. Bradley , Blackwell Jonathan E. , Ou Jao J. , Shinde Patil Vivek R. , Goetz Douglas J.
Published: August 2002

In a variety of disease settings the expression of the endothelial selectins E‐ and P‐selectin appears to be increased. This feature makes these molecules attractive targets around which to design directed drug‐delivery schemes. One possible approach for achieving such delivery is to use polymeric biodegradable microspheres bearing a humanized monoclonal antibody (MAb) for E‐ and P‐selectin, MAb HuEP5C7.g2. Perhaps the simplest technique for “coupling” HuEP5C7.g2 to the microspheres is via nonspecific adsorption. Previous studies suggest, however, that the adsorption of proteins onto microspheres fabricated in the presence of a stabilizer such as poly(vinyl alcohol) (PVA) is limited. It is unclear to what extent this limited level of adsorbed HuEP5C7.g2 would be able to support adhesion to E‐ and P‐selectin under flow conditions. To explore this issue, we prepared microspheres from the biodegradable polymer, poly(ϵ‐caprolactone) (PCL), using a single emulsion process and PVA as a stabilizer. We then incubated the PCL microspheres with HuEP5C7.g2 and studied the adhesion of the resulting HuEP5C7.g2 microspheres to E‐ and P‐selectin under in vitro flow conditions. We found that the HuEP5C7.g2 PCL microspheres exhibit specific adhesion to Chinese hamster ovary cells stably expressing P‐selectin (CHO‐P) and 4‐h IL‐1β–activated human umbilical vein endothelial cells (HUVEC). In contrast, HuEP5C7.g2 PCL microspheres exhibit little adhesion to parental CHO cells or unactivated HUVEC. The attachment efficiency to the selectin substrates was quite low, with appreciable attachment occurring only at low shear (0.3 dyn/cm2). Other supporting data strongly suggest that the limited attachment efficiency is due to a low level of HuEP5C7.g2 adsorbed to the PCL microspheres. Although the attachment was limited, a significant percentage of the HuEP5C7.g2 PCL microspheres were able to remain adherent at relatively high shear (8 dyn/cm2). Combined, our data suggest that HuEP5C7.g2 PCL microspheres exhibit selective limited adhesion to cellular substrate expressing E‐ and P‐selectin. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 73: 500–509, 2001.
A chemostat culture as a tool for the improvement of a recombinant E. coli strain over‐producing penicillin G acylase
Authors: Marešová H. , Štěpánek V. , Kyslík P.
Published: August 2002

The recombinant strain RE3(pKA18) of Escherichia coli constitutively overproduces penicillin G acylase (PGA) from plasmid‐borne gene pga. The host strain RE3 bears the same pga gene on its chromosome, the expression of which is controlled by the natural mechanism of induction with phenylacetic acid (PA). To evaluate the maximum biosynthetic capacity for PGA, induction of the chromosomal pga by PA was studied in a culture of the recombinant strain. PGA production by batch cultures of RE3(pKA18) and RE3 showed a different response to the addition of PA to the medium: while an addition of PA induces PGA in a culture of strain RE3 as expected, in recombinant cells it lowers the specific activity of PGA and a large amount of PGA is released into the culture medium. To improve the PGA production, the strain RE3(pKA18) was cultured in a carbon‐limited chemostat and subjected to selection pressure in a medium supplemented with phenylacetic acid amide (PAA). Phenylacetic acid amide served as a source of nitrogen, an inducer of PGA and a factor exerting positive selection pressure on the maintenance of the recombinant plasmid. After 130 generations of growth in the presence of the inducer, no recombinant strain with constitutive expression of the chromosomal gene pga was detected in the prevailing P+ subpopulation in the chemostat. Shake‐flask experiments with the parent recombinant strain RE3(pKA18), host strain RE3, chemostat evolvant ERE3(epKA18), the cured host ERE3 alone, and its derivative after retransformation with ancestral plasmid ERE3(pKA18) showed that inactivation of the plasmid‐borne pga by a frame‐shift mutation (plasmid epKA18) occurred in the plasmid‐bearing subpopulation accumulated in the chemostat. Marked adaptive changes evolved in the host ERE3 during a 130 generation culture: (1) the specific growth rate of the host increased by 30% in a medium without PA, (2) the copy number of plasmids pKA18 and epKA18 in the host cultured in PA‐free medium dropped by about 40%, and (3) the leakage of PGA from the cell in the presence of PA found in strain RE3(pKA18) was not observed in strain ERE3(pKA18). This new recombinant strain with modified traits was constructed by means of retransformation of the evolved host ERE3 with ancestral plasmid pKA18. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 46–52, 2001.
Resolution of enantiomers of DL‐amino acids on silica gel plates impregnated with optically pure (—)‐quinine
Authors: Bhushan R. , Arora Meenakshi
Published: August 2002

TLC resolution of enantiomers from racemic amino acids was achieved on silica gel plates impregnated with optically pure (—)‐quinine. The successful solvent systems were butanol–chloroform–acetic acid (3:7:5, v/v) for DL‐methionine; 6:8:4, v/v for alanine; 10:1:4; v/v for threonine; and ethyl acetate–carbon tetrachloride–propionic acid (10.5:6.5:3.5, v/v) for valine. Minimum detection limits were found to be different for each of the amino acid, ranging between 0.9 and 3.7 µg. The effects of concentration of impregnating reagent, temperature and pH on resolution of enantiomers have been studied in details. Copyright © 2001 John Wiley & Sons, Ltd.
The application of an action system model to destructive behaviour: the examples of arson and terrorism
Authors: Fritzon Katarina , Canter David , Wilton Zoe
Published: August 2002

This article argues that criminal and deviant behaviour can be productively viewed through an action system framework. The idea is developed by considering two forms of destructive behaviour: arson and barricade–hostage terrorist incidents. Two studies are presented. The first study tests the hypothesis that different forms of arson will reflect the four dominant states that an action system can take; integrative, expressive, conservative, and adaptive. A smallest space analysis was performed on 46 variables describing 230 cases of arson and the results identified the four themes of action system functioning. An examination of the personal characteristics of the arsonists also produced four variable groupings and a combined analysis of the four action scales and four characteristics scales also supported the structural hypothesis of the action system model. The second study applied the action system model to the study acts of terrorist barricade–hostage incidents. A smallest space analysis of 44 variables coded from 41 incidents again revealed four distinct forms of activity, which were psychologically similar to the four modes of arson identified in study one. Overall, these two studies provide support for the appropriateness of the action system framework as a way of classifying different forms of deviant behaviour. Copyright © 2001 John Wiley & Sons, Ltd.
Anatomic and clinical correlations of the lenticulostriate arteries
Authors: Marinković S. , Gibo H. , Milisavljević M. , Ćetković M.
Published: August 2002

The authors examined the lenticulostriate (perforating) arteries in the vascular casts of 48 middle cerebral arteries (MCA), as well as in the MRI or CT scans of 32 patients with cerebral infarcts in the MCA territory. The lenticulostriate arteries ranged between two and 12 in number, and from 80 μm to 1,400 μm in size. They originated from the main trunk, terminal trunks, bifurcation site, and/or leptomeningeal branches of the MCA, either separately or from common trunks (70.8%). The extreme variations of the supplying region of the perforators were noted in seven anatomic specimens. In addition to the basal ganglia, the genu, and the anterior limb of the internal capsule, the lenticulostriate arteries seemed to supply only the rostral portion of the superior part of the posterior limb of the capsule. The patients presented with occlusion of all the lenticulostriate arteries, individual arteries, or only their twigs. Complete occlusion of these arteries resulted in a huge central hemispheric infarct. Occlusion of an individual artery most often caused a large ganglionic‐capsular infarct. The authors concluded that the lacunar infarcts usually follow occlusion of a terminal or a side branch of the lenticulostriate arteries. Clin. Anat. 14:190–195, 2001. © 2001 Wiley‐Liss, Inc.
Community psychology: should there be a European perspective?
Authors: Francescato Donata , Tomai Manuela
Published: August 2002

In this era of globalisation community psychologists have to examine how globalisation patterns interact with local cultural norms, to find tools to promote a sense of community that fits a particular context. We cannot therefore acritically adopt for many European contexts, community psychology concepts and intervention strategies geared to USA values. The paper argues for the need to develop a European perspective in Community Psychology, built more on the European tradition of political concern for promoting social capital, besides an individual's freedom and autonomy. The paper attempts to identity some of the main differences that have emerged in the last decades between USA and European approaches to community psychology. It also describes two empowering tools, which integrate traditional and post modern views of science: community profiling and multidimensional organisational analysis, that have been used by European community psychologists to rebuild social capital in organisations and local communities. Copyright © 2001 John Wiley & Sons, Ltd.
Genetic polymorphism of cytochrome P450 2E1 in the Turkish population
Authors: Ömer Beyhan , Özbek Uğur , Akköse Ayfer , Kiliç Gamze
Published: August 2002

CYP 2E1 is involved in metabolic activation of carcinogenic N‐nitrosamines, benzene, urethane and other low molecular weight compounds. CYP2E1 gene is present in the population in various polymorphic forms. We detected the RFLP of the human CYP2E1 gene with the restriction endonuclease PstI, RsaI and DraI in a group of 153 Turkish individuals. According to the results of the PstI/RsaI analysis, 96.07% of the subjects were of the c1/c1 genotype, and 3.93% were of the c1/c2 genotype. In the DraI RFLP analysis, 84.30% DD genotype, 15.03% CD genotype and 0.66% CC genotype were determined. The data obtained may be useful in epidemiological studies of the influence of CYP2E1 polymorphism on carcinogenesis. Copyright © 2001 John Wiley & Sons, Ltd.
Same‐day transradial outpatient stenting with a 6‐hr course of glycoprotein IIb/IIIa receptor blockade: A feasibility study
Authors: Gilchrist Ian C. , Nickolaus Michelle J. , Momplaisir Thierry
Published: August 2002

The aim of the study was to explore the feasibility of same‐day outpatient stent placement using a short course of intravenous antiplatelet therapy. Patients (n = 26) had stent placement and 6 hr of eptifibatide therapy. Demographics, procedural information, CPK data, and length of stay were recorded along with postdischarge outcomes. Twenty‐one men and five women with median age of 60 years (49, 69) underwent transradial stenting. Baseline characteristics included diabetes 62%, hyperlipidemia 77%, prior coronary bypass surgery 19%, and unstable angina 35%. There were no CPK elevations (> 2 × normal) or ECG changes. Discharge occurred after 6.5 hr (5.8, 7.0). Neither vascular site complications nor readmission for procedure‐related problems occurred. One patient later expressed concerns about discharge education. Outpatient stent placement with 6‐hr infusion of GP IIb/IIIa inhibitor appears feasible and efficient in select patients. There may be challenges to meet with regard to patient education. Further studies with larger populations are needed to evaluate and optimize this approach. Cathet Cardiovasc Intervent 2002;56:10–13. © 2002 Wiley‐Liss, Inc.
Coronary angiography using 4 Fr catheters with acisted power injection: A randomized comparison to 6 Fr manual technique and early ambulation
Authors: Khoukaz Souheil , Kern Morton J. , Bitar Saad R. , Azrak Elie , Eisenhauer Michael , Wolford Thomas , El‐Shafei Amr
Published: August 2002

Coronary angiography using 4 Fr catheters may reduce access site complications, promote better utilization of outpatient facilities, but at a cost of suboptimal image quality. To determine whether 4 Fr diagnostic angiography with power injection (Acist, Minneapolis, MN) was equivalent to 6 Fr manual technique, 101 unselected patients were randomized to transfemoral coronary angiography with 4 or 6 Fr catheters. Procedural characteristics, angiographic quality scores, and results of 90 min ambulation were analyzed. Coronary angiographic quality scores using 4 Fr and 6 Fr catheters were equivalent (left coronary artery 4.73 ± 0.6 vs. 4.80 ± 0.65, P = 0.28; right coronary artery 4.98 ± 90.13 vs. 4.97 ± 0.16, P = 0.48). However, 4 Fr left ventriculographic image score was lower (4.53 ± 0.68 vs. 4.83 ± 0.42, P = 0.0002), attributed, in part, to a smaller injected contrast volume (32 ± 11 vs. 37 ± 4 mL, P = 0.001). The total study contrast volume was significantly less in the 4 Fr group (119 ± 35 vs. 159 ± 52 mL, P = 0.001). Complications related to early ambulation at 90 min were similar and minimal in both groups. Compared to 6 Fr manual contrast injection technique, diagnostic angiography through 4 Fr catheters with power contrast injection resulted in equivalent coronary angiographic image quality, slightly reduced but diagnostic left ventricular image quality, and significantly less contrast volume. Four Fr angiography facilitates early ambulation without compromising safety and image quality. Cathet Cardiovasc Intervent 2001;52:393–398. © 2001 Wiley‐Liss, Inc.
Use of ICHOR‐platelet works to assess platelet function in patients treated with GP IIb/IIIa inhibitors
Authors: Lakkis Nasser M. , George Sima , Thomas Elson , Ali Mohamad , Guyer Kirk , Carville David
Published: August 2002

Platelet glycoprotein GP IIb/IIIa inhibitors have been recently approved for use in treating patients with acute coronary syndromes and those undergoing PCI. The purpose of this study was to assess the feasibility of using a new device, the ICHOR platelet works, to detect platelet inhibition in patients undergoing PCI and treated with abciximab or tirofiban. The study was conducted at Baylor College of Medicine, Houston, Texas. Thirty patients undergoing PCI and treated with abciximab (n = 10) or tirofiban (n = 20) are included. Blood samples were obtained before, at 30 min, at 4 hr, and at 12 hr after starting the GP IIb/IIIa inhibitors and 2 hr after discontinuation. Baseline studies revealed > 95% platelet aggregability in all patients after exposure to ADP (20 μM). After starting tirofiban, 82%, 83%, and 82% of platelets were inhibited at 30 min, 4 hr, and 12 hr. Platelet inhibition decreased to 43% 2 hr after discontinuation of tirofiban. Similarly, ICHOR platelet works detected 91%, 92%, and 85% platelet inhibition at 30 min, 4 hr, and 12 hr after starting abciximab, respectively. Platelet inhibition decreased to 73% 2 hr after discontinuation. The ICHOR platelet works is a promising, simple, and rapid bedside method that may have clinical utility in assessing platelet inhibition in patients treated with GP IIb/IIIa inhibitors. Cathet Cardiovasc Intervent 2001;53:346–351. © 2001 Wiley‐Liss, Inc.
Stenting of coarctation of the aorta
Authors: Cheatham John P.
Published: August 2002

The purpose of the present study was to assess the usefulness of balloon expandable Palmaz intravascular stents in the transcatheter treatment of children and young adults with native and recurrent coarctation of the aorta, and to develop an improved intravascular stent and balloon delivery catheter specifically developed for vascular obstructions associated with congenital heart disease, including coarctation of the aorta. Twenty‐one patients, 8 native and 13 recurrent coarctations, were successfully treated with the Palmaz stent. However, limitations and disadvantages in stent design and the single balloon delivery system were uncovered. Therefore, the NuMED CP stent and BIB delivery catheter were developed and used to treat 25 patients with native (17) and recurrent (8) coarctation successfully. Improvements in stent design and long‐term follow‐up using three‐dimensional spiral CT scan will be helpful in determining the role of transcatheter stent therapy for native and recurrent coarctation of the aorta. Cathet Cardiovasc Intervent 2001;54:112–125. © 2001 Wiley‐Liss, Inc.
The low down on e‐science and grids for biology
Authors: Goble Carole
Published: August 2002

The Grid is touted as a next generation Internet/Web, designed primarily
to support e‐Science. I hope to shed some light on what the Grid is,
its purpose, and its potential impact on scientific practice in biology. The
key message is that biologists are already primarily working in a manner that
the Grid is intended to support. However, to ensure that the Grid's
good intentions are appropriate and fulfilled in practice, biologists must
become engaged in the process of its development. Copyright © 2001 John Wiley & Sons, Ltd.
Website Review: structural genomics on the web
Authors: Wixon Jo
Published: August 2002

In this review we provide a brief guide to some of the resources and databases that can be used to locate information and aid research in the growing field of structural genomics. The review will provide examples, for less experienced users, of what can be achieved using a selection of the available sites. We hope that this will encourage you to use these sites to their full potential and whet your appetite to search for other related sites. Copyright © 2001 John Wiley & Sons, Ltd.
Combining genomics, metabolome analysis, and biochemical modelling to understand metabolic networks
Authors: Fiehn Oliver
Published: August 2002

Now that complete genome sequences are available for a variety of organisms, the elucidation of gene functions involved in metabolism necessarily includes a better understanding of cellular responses upon mutations on all levels of gene products, mRNA, proteins, and metabolites. Such progress is essential since the observable properties of organisms – the phenotypes – are produced by the genotype in juxtaposition with the environment. Whereas much has been done to make mRNA and protein profiling possible, considerably less effort has been put into profiling the end products of gene expression, metabolites. To date, analytical approaches have been aimed primarily at the accurate quantification of a number of pre‐defined target metabolites, or at producing fingerprints of metabolic changes without individually determining metabolite identities. Neither of these approaches allows the formation of an in‐depth understanding of the biochemical behaviour within metabolic networks. Yet, by carefully choosing protocols for sample preparation and analytical techniques, a number of chemically different classes of compounds can be quantified simultaneously to enable such understanding. In this review, the terms describing various metabolite‐oriented approaches are given, and the differences among these approaches are outlined. Metabolite target analysis, metabolite profiling, metabolomics, and metabolic fingerprinting are considered. For each approach, a number of examples are given, and potential applications are discussed. Copyright © 2001 John Wiley & Sons, Ltd.
Meeting Review: The Second European Conference on Zebrafish Genetics and Development
Authors: Feldman Benjamin
Published: August 2002

Meeting Review of the Second European Conference on Zebrafish Genetics and Development, held at University College London, England, April 19‐22, 2001.
SAND, a new protein family: from nucleic acid to protein structure and function prediction
Authors: Cottage Amanda , Edwards Yvonne J. K. , Elgar Greg
Published: August 2002

As a result of genome, EST and cDNA sequencing projects, there are huge numbers of predicted and/or partially characterised protein sequences compared with a relatively small number of proteins with experimentally determined function and structure. Thus, there is a considerable attention focused on the accurate prediction of gene function and structure from sequence by using bioinformatics. In the course of our analysis of genomic sequence from Fugu rubripes, we identified a novel gene, SAND, with significant sequence identity to hypothetical proteins predicted in Saccharomyces cerevisiae, Schizosaccharomyces pombe, Caenorhabditis elegans, a Drosophila melanogaster gene, and mouse and human cDNAs. Here we identify a further SAND homologue in human and Arabidopsis thaliana by use of standard computational tools. We describe the genomic organisation of SAND in these evolutionarily divergent species and identify sequence homologues from EST database searches confirming the expression of SAND in over 20 different eukaryotes. We confirm the expression of two different SAND paralogues in mammals and determine expression of one SAND in other vertebrates and eukaryotes. Furthermore, we predict structural properties of SAND, and characterise conserved sequence motifs in this protein family. Copyright © 2001 John Wiley & Sons, Ltd.
Enantioselective carbon‐‐hydrogen insertion is an effective and efficient methodology for the synthesis of (r)‐(‐)‐baclofen
Authors: Doyle Michael P. , Hu Wenhao
Published: August 2002

A highly enantioselective methodology for the synthesis of the GABAB receptor agonist (R)‐(‐)‐baclofen is described. This synthesis begins with p‐chlorophenethyl alcohol and involves a catalytic carbon‐hydrogen insertion reaction of a chiral dirhodium(II) carboxamidate with the corresponding diazoacetate (81% yield, 95% ee). Subsequent steps convert the intermediate γ‐lactone to (R)‐ (‐)‐baclofen in a 60% overall yield. The amount of catalyst required for the C–H insertion transformation is only 0.5 mol%. Chirality 14:169–172, 2002.© 2002 Wiley‐Liss, Inc.
Asymmetric catalytic ene‐cyclization approach to 2–fluoro‐19‐nor‐1,25‐dihydroxyvitamin D3 A‐ring analog with significant transactivation activity
Authors: Mikami Koichi , Ohba Shiho , Ohmura Hirofumi , Kubodera Noboru , Nakagawa Kimie , Okano Toshio
Published: August 2002

1α,25‐Dihydroxyvitamin D3 (1α,25(OH)2D3) has been shown to modulate not only proliferation and differentiation, but also apoptosis in malignant cells, indicating that it could be useful for the treatment of cancer and psoriasis. However, little information has been available on the binding conformation of the 1α,25(OH)2D3 molecule and its analogs with the vitamin D receptor (VDR). Therefore, we synthesized 2α‐fluorinated A‐ring analogs of 19‐nor‐1α,25(OH)2D3 in order to investigate the VDR‐binding conformation of the A‐rings on the basis of the 19F NMR analysis. The 2α–fluoro‐19‐nor‐1α,25‐dihydroxyvitamin D3 A‐ring analog thus synthesized via a asymmetric catalytic carbonyl‐ene cyclization, shows significant activity in transactivation. Chirality 13:366–371, 2001. © 2001 Wiley‐Liss, Inc.
Geometric Clutch model version 3: The role of the inner and outer arm dyneins in the ciliary beat
Authors: Lindemann Charles B.
Published: August 2002

The Geometric Clutch model of ciliary and flagellar beating uses the transverse force (t‐force) that develops between the outer doublets of the axoneme as the regulator for activating and deactivating the dynein motors and organizing the flagellar beat. The version of the model described here adds detail to the formulations used in the two previous versions as follows: (1) In place of two opposing sets of dyneins, the new model has four sets of dyneins, corresponding to two sets on each side of the axoneme acting in series. (2) The four sets of dyneins are each subdivided into two ranks representing inner and outer arm dyneins. (3) The force produced by each dynein is governed by a force‐velocity relationship that is independently specified for the inner and outer arms. Consistent with the original model, the new version of the Geometric Clutch model can simulate both the effective and recovery stroke phases of the ciliary beat using a single uniform algorithm. In addition, the new version can operate with the outer arms disabled. Under this condition, the simulation exhibits a beat pattern similar to the original but the beat frequency is reduced to approximately one third. These results are contingent on using force‐velocity relationships for the inner and outer arms similar to those described by Brokaw [1999: Cell Motil. Cytoskeleton 42:134–148], where the inner arms contribute most of the driving force at low shear velocities. This constitutes the first examination of the effects of the force‐velocity characteristics of dynein on a cilia‐like beat in a theoretical framework. Cell Motil. Cytoskeleton 52:242–254, 2002. © 2002 Wiley‐Liss, Inc.
Enhanced expression of telomerase activity in thymoma and thymic carcinoma tissues
Authors: Watanabe Masazumi , Yu Shan‐kang , Sawafuji Makoto , Kawamura Masafumi , Horinouchi Hirohisa , Mukai Makio , Kobayashi Koichi
Published: August 2002

BACKGROUND

Telomerase is a nucleoprotein complex that caps the physical termini of all eukaryotic chromosomes. Because most malignant cells and reproductive cells have telomerase activity, which elongates telomeric DNA, telomerase may play important roles in unlimited cell division acquisition of the malignant phenotype. The current study examined the relation of telomerase activity in thymoma and thymic carcinoma with the clinicopathologic features of these lesions.

METHODS

Tissue specimens were surgically resected from patients with thymoma and thymic carcinoma. Telomerase activity was evaluated according to a modified telomeric repeat amplification protocol assay. Paraffin sections of tumor were immunostained by MIC2 antibody, a marker of immature T cells.

RESULTS

Telomerase activity was detected in all thymic epithelial tumors. The activity (mean ± SD; unit per μg protein) in thymoma (n = 17) was significantly higher than that in thymic carcinoma (n = 7) (431.8 ± 400.1 vs. 68.8 ± 39.8; P < 0.01). Telomerase activities in thymoma and thymic carcinoma were significantly higher than that in primary lung adenocarcinoma (33.5 ± 39.2, n = 47), studied as a control (P < 0.01). In patients with thymoma, telomerase activity did not correlate with tumor stage according to Masaoka classification (P = 0.776). In patients with thymic carcinoma, however, telomerase activity positively correlated with tumor stage (P = 0.02). In thymoma, telomerase activity positively correlated with the ratio of induced lymphocytes according to Rosai's classification (P = 0.045). MIC2‐positive lymphocytes were identified in all cases of thymoma (n = 12). In contrast, lymphocytes infiltrating thymic carcinoma did not react with MIC2.

CONCLUSIONS

In thymoma, telomerase activity reflects the presence of immature T‐cell lymphocytes in tumor tissue rather than tumor stage or malignant phenotype. In thymic carcinoma, telomerase activity derived directly from cancer cells may relate to tumor stage. Cancer 2002;94:240–4. © 2002 American Cancer Society.
Testosterone recovery following prolonged adjuvant androgen ablation for prostate carcinoma
Authors: Pickles Tom , Agranovich Alex , Berthelet Eric , Duncan Graeme G. , Keyes Mira , Kwan Winkle , McKenzie Michael R. , Morris W. James ,   the British Columbia Cancer Agency Prostate Cohort Outcomes Initiative
Published: August 2002

BACKGROUND

This study was conducted to describe the rate and completeness of the recovery of testosterone production following prolonged temporary androgen ablative therapy in men with prostate carcinoma undergoing curative radiation therapy.

METHODS

Two‐hundred and sixty‐seven men treated with between 3 months and 3 years of adjuvant androgen ablation (AA) were followed at 6‐month intervals following cessation of their androgen deprivation therapy. A comparative group of 518 men not undergoing AA were also followed.

RESULTS

Drugs used included low dose cyproterone/stilboestrol (CPA/DES) in combination (56%) and 1 month depot (18%) and 3 month depot (25%) leutinizing hormone releasing hormone agonist (LHRHa). Seventy‐nine percent of men in the current study recovered normal testosterone levels (10nmol/L), and 93% recovered levels of at least 5nmol/L. In comparison, men who had never received androgen ablative therapy showed a fall of testosterone, with 17% having sub‐normal levels after 3 years. Median time to testosterone recovery was 10 months. Factors associated on multivariate analysis with delayed testosterone recovery included advanced age (P = 0.008), low pre‐therapy testosterone (P = 0.04), and the use of 3 month LHRHa preparations as compared with CPA/DES (P = 0.002) or 1 month LHRHa preparations (P = 0.015). The duration of drug use was not significantly associated with time to testosterone recovery.

CONCLUSIONS

Long‐acting LHRHa preparations appear to have a more prolonged action than previously supposed. Most men treated for up to 2 years recover normal testosterone levels after cessation of adjuvant androgen ablation, and the limited data available in the current study on patients treated for 3 years also suggests most will recover. Cancer 2002;94:362–7. © 2002 American Cancer Society.
The APC/β‐catenin pathway in ulcerative colitis–related colorectal carcinomas
Authors: Aust Daniela E. , Terdiman Jonathan P. , Willenbucher Robert F. , Chang Cornell G. , Molinaro‐Clark Annette , Baretton Gustavo B. , Loehrs Udo , Waldman Frederic M.
Published: August 2002

BACKGROUND

Although the APC/β‐catenin pathway is known to play a crucial role in sporadic colorectal carcinogenesis, its influence on ulcerative colitis (UC)–related neoplastic progression is unknown. To elucidate the role of the APC‐/β‐catenin pathway in UC‐related carcinogenesis, the authors identified APC and β‐catenin mutations in a set of UC‐related and sporadic colorectal carcinomas.

METHODS

The mutational cluster region of APC (codon 1267 to 1529) and exon 3 of the β‐catenin were directly sequenced.

RESULTS

Only 1 of 30 UC‐related tumors (3%) showed an APC mutation whereas 11 of the 42 sporadic carcinomas (26%) had mutations within the mutational cluster region. Within the sporadic carcinoma group, only 8% of the right‐sided carcinomas showed APC mutations whereas 50% of the left‐sided carcinomas had mutations within the mutational cluster region. None of the tumors in either group showed a β‐catenin mutation.

CONCLUSIONS

Mutations of the APC and β‐catenin are rare in UC‐related tumors. These genes may be altered because of mutations outside the regions studied, or by epigenetic silencing. Alternatively, other proteins involved in the APC/β‐catenin signaling cascade may be altered, or this pathway may be involved infrequently in UC‐related carcinogenesis. The significant difference in frequency of APC mutations between right‐ and left‐sided sporadic tumors suggests different molecular pathways in these two tumor sites. Cancer 2002;94:1421–7. © 2002 American Cancer Society.

DOI 10.1002/cncr.10334
Detection of recurrent or persistent nasopharyngeal carcinomas after radiotherapy with technetium‐99m methoxyisobutylisonitrile single photon emission computed tomography and computed tomography
Authors: Kao Chia‐Hung , Shiau Yu‐Chien , Shen Yeh‐You , Yen Ruoh‐Fang
Published: August 2002

BACKGROUND

The diagnostic accuracy of technetium‐99m methoxyisobutylisonitrile (Tc‐MIBI) single photon emission computed tomography (SPECT) and computed tomography (CT) of the head and neck for differentiating recurrent or residual nasopharyngeal carcinomas (NPC) from benign lesions after radiotherapy was compared with positron emission tomography (PET) with 18‐fluoro‐2‐deoxyglucose (FDG).

METHODS

Thirty six NPC patients underwent head and neck CT, Tc‐MIBI SPECT, and FDG‐PET four months after radiotherapy to differentiate recurrent or residual NPC from benign lesions. Histopathologic examinations of nasopharyngeal biopsies were performed for all 36 patients. No patients had multiple foci of NPC.

RESULTS

Based on biopsy results, the sensitivity, specificity, and accuracy of CT for differentiating recurrent or residual NPC from benign lesions were 73%, 88%, and 83%, respectively. The sensitivity, specificity, and accuracy of Tc‐MIBI SPECT were 73%, 96%, and 89%, respectively. The sensitivity, specificity, and accuracy of FDG‐PET were 100%, 96%, and 97%, respectively. Combination CT and Tc‐MIBI SPECT for 28 NPC patients with congruent results showed the same sensitivity, specificity, and accuracy of 100%, 96%, and 96%, respectively, as FDG‐PET for differentiating recurrent or residual NPC from benign lesions. In eight patients with incongruent results between CT and Tc‐MIBI SPECT, FDG‐PET correctly differentiated two benign lesions and six recurrent or residual NPCs.

CONCLUSIONS

In detecting recurrent or residual NPC, FDG‐PET is the best tool. However, combined use of CT and Tc‐MIBI SPECT can result in the same accuracy as FDG‐PET. Cancer 2002;94:1981–6. © 2002 American Cancer Society.

DOI 10.1002/cncr.10400
Intraoperative evaluation of sentinel lymph nodes for metastatic melanoma by imprint cytology
Authors: Creager Andrew J. , Shiver Stephen A. , Shen Perry , Geisinger Kim R. , Levine Edward A.
Published: August 2002

BACKGROUND

Sentinel lymph node (SLN) biopsy has revolutionized lymph node staging in patients with malignant melanoma. Intraoperative evaluation is a new addition to the SLN procedure that allows for a one‐step regional lymph node dissection to be performed when the SLN biopsy findings are positive. To date, several studies have evaluated the use of intraoperative frozen sectioning to evaluate the SLN in patients with melanoma. The literature pertaining to the use of intraoperative imprint cytology (IIC) to evaluate the SLN in melanoma patients is scant and to the authors' knowledge studies published to date are relatively small. The purpose of the current study was to evaluate the utility of IIC in patients undergoing SLN for melanoma.

METHODS

A total of 235 SLN biopsies from 93 patients with malignant melanoma and 3 patients with atypical Spitz nevi were examined by IIC after SLN biopsy using a double indicator technique. The SLNs were bisected and a pair of imprints were made from each half. One imprint from each half was stained with hematoxylin and eosin (H & E) whereas its counterpart was stained with Diff‐Quik. Paraffin‐embedded permanent sections were examined using multiple H & E stained sections from the SLNs in conjunction with immunohistochemical staining for S‐100 and HMB‐45 proteins.

RESULTS

A total of 235 SLNs were excised from 93 patients (2.5 SLNs per patient). On a per patient basis, metastases were identified in 21 patients (23%) on permanent section evaluation. Of these 21 patients, 8 were detected by IIC (sensitivity of 38%). The negative predictive value was 85%. No false‐positive results were identified (specificity of 100%). The positive predictive value was 100%. The overall accuracy of the intraoperative evaluation was 86%. Patients found to have positive SLNs by IIC went on to undergo lymphadenectomy under the same anesthetic.

CONCLUSIONS

The sensitivity and specificity of IIC are similar to those of intraoperative frozen‐section evaluation. Therefore, IIC appears to be a viable alternative to frozen sectioning when intraoperative evaluation is required. IIC evaluation of SLN makes a single surgical procedure possible for patients with malignant melanoma who are undergoing SLN. Cancer 2002;94:3016–22. © 2002 American Cancer Society.

DOI 10.1002/cncr.10512
Racial/ethnic differences in survival rates in a population‐based series of men with breast carcinoma†
Authors: O'Malley Cynthia D. , Prehn Angela W. , Shema Sarah J. , Glaser Sally L.
Published: August 2002

BACKGROUND

A rare occurrence, about 1500 men in the United States develop breast carcinoma each year. Little is known about survival patterns at the population level, particularly about racial/ethnic variation.

METHODS

Using data from the Surveillance, Epidemiology, and End Results Program, we examined survival rates in 1979 men diagnosed with primary invasive breast carcinoma between 1973 and 1997. Race was defined as non‐Hispanic white, non‐Hispanic black, and other race/ethnicity (predominantly Asian/Pacific Islander and Hispanic). The two outcomes were all‐cause and breast carcinoma‐ specific mortality. Survival curves were drawn using Kaplan–Meier estimates and Cox regression was used to estimate the risk of death with hazard ratios and 95% confidence intervals. For both outcomes, the racial/ethnic survival curves differed significantly when the log rank test was used. Therefore, separate models were run for each racial/ethnic group. Covariates included age, stage, histology, surgery, radiation therapy, and year of diagnosis. Estrogen and progesterone receptor status were available for 616 men.

RESULTS

Survival rates differed significantly by race/ethnicity. Overall, 5‐year survival rates were 66% for whites, 57% for blacks, and 75% for men of other race/ethnicity. Blacks presented with more advanced disease. By stage, whites and blacks had worse survival rates compared with men of other race/ethnicity. The effects of prognostic factors such as age, surgery type, and radiation were similar, but not always significant, for all groups. Diagnosis year and estrogen receptor status did not affect survival.

CONCLUSIONS

Survival following male breast carcinoma differed by race/ethnicity, whereas the prognostic factors associated with survival were similar. Cancer 2002;94:2836–43. Published 2002 American Cancer Society.

DOI 10.1002/cncr.10521
Role of retinal afferents in regulating growth and shape of the lateral geniculate nucleus
Authors: Williams Adrian L. , Reese Benjamin E. , Jeffery Glen
Published: August 2002

Segregated binocular maps in the dorsal lateral geniculate nucleus (LGN) develop from a stage where they initially overlap. Sophisticated computational models have been used to describe the dynamics of three‐dimensional LGN shape changes that play a role in segregation. These models have revealed specific nuclear growth vectors associated with the process of ocular segregation in the LGN (Williams and Jeffery [2001] J Comp Neurol 430:332–342.). In this study, we use similar techniques to determine whether retinal innervation contributes to the dynamics of shape maturation in the ferret LGN. In this animal, 90% of the retinal innervation of the mature LGN comes from the contralateral eye. If one eye is removed before segregation, the projection from the remaining eye remains diffuse and nuclear growth is stunted. Here, we quantify this effect and show that removing the contralateral projection before segregation has a profound impact on LGN size but changes its ultimate shape by only 12%. The impact on shape on the other side of the brain where the ipsilateral projection is removed, which accounts for only 10% of its innervation in maturity, is less than 2%. Hence, retinal innervation plays a minor role in determining mature LGN shape. Although in both hemispheres, the ultimate shape of the nucleus is close to normal, removal of the larger projection disrupts normal growth vectors, with almost none being present in the 5 days after enucleation, when the normal nucleus expands markedly. Hence, the effect of enucleation is to delay shape maturation. Growth vectors absent after removal of the smaller projection are mainly confined to those in what would be the binocular region. J. Comp. Neurol. 445:269–277, 2002. © 2002 Wiley‐Liss, Inc.
High‐resolution localization of clathrin assembly protein AP180 in the presynaptic terminals of mammalian neurons
Authors: Yao Pamela J. , Coleman Paul D. , Calkins David J.
Published: August 2002

Synaptic vesicles (SVs) assemble at the presynaptic compartment through a clathrin‐dependent mechanism that involves one or more assembly proteins (APs). The assembly protein AP180 is especially efficient at facilitating clathrin cage formation, but its precise ultrastructural localization in neurons is unknown. Using immunoelectron microscopy, we demonstrate the presynaptic localization of AP180 in axon terminals of rat cerebellar neurons. In contrast, the assembly protein AP2 was associated with both the presynaptic plasma membrane and the cytosolic side of the membrane at postsynaptic and extrasynaptic sites. Furthermore, ultrastructural analysis of primate retina showed that AP180 immunoreactivity was preferentially and highly enriched at ribbon synapses, where glutamate is released tonically at high levels and rapid vesicle turnover is essential. To maintain functional synaptic transmission, neurotransmitter‐filled SVs must be readily available, and this requires proper reassembly of new vesicles. The expression of AP180, in addition to AP‐2, in the clathrin‐mediated endocytic pathway might add another level of control to SV reformation for efficient assembly of clathrin, effectively controlling the size of assembled vesicles and faithfully recovering SV‐specific components. J. Comp. Neurol. 447:152–162, 2002. © 2002 Wiley‐Liss, Inc.
On counting and counting errors
Authors: Guillery R.W.
Published: August 2002

Counting objects in histological sections is often a necessary, sometimes an unexpected part of a research project. The recent literature shows that the subject of counting is of particular interest to readers of the Journal of Comparative Neurology but that it is also contentious and difficult. Even a brief review of past issues of the Journal shows that there are many misconceptions about counting and that there remain issues that have received little or no attention. Counts are subject to many errors. Some reports include readily recognizable errors, others fail to include all of the information that is needed for an evaluation of their accuracy. This review is above all a plea for adequate information about the methods used for counts in all publications. It serves to help those who are new to quantitative methods in histology; it considers some of the basic issues arising for anyone undertaking counts, or reviewing manuscripts that include counts. In particular, it considers recently introduced or re‐introduced counting methods that depend on accurate measures along the axis perpendicular to the plane of the sections, and looks at the difficulties inherent in these measures. J. Comp. Neurol. 447:1–7, 2002. © 2002 Wiley‐Liss, Inc.
Differential localization of high‐ and low‐molecular‐weight variants of microtubule‐associated protein 2 in the developing rat telencephalon
Authors: Fujimori Kazuhiro , Takauji Rumiko , Tamamaki Nobuaki
Published: August 2002

Microtubule‐associated protein 2 (MAP2) occurs in developing mammalian neuronal tissue as both high‐ and low‐molecular‐weight forms with temporally regulated expression. We studied the MAP2 expression in the developing rat telencephalon with monoclonal antibodies that recognized both the high‐ and low‐molecular‐weight forms of MAP2 variants or that specifically recognized high‐molecular‐weight forms of MAP2 variants. Differences in the staining patterns of these antibodies reflected differences in the distribution of the high‐ and low‐molecular‐weight MAP2s. The immunoreactive sites of high‐ and low‐molecular‐weight MAP2 had a more widespread distribution in the embryonic telencephalon than those of high‐molecular‐weight MAP2. Many bipolar cells in the ganglionic eminence (GE) and in the intermediate zone (IZ) of the neocortex showed low‐molecular‐weight MAP2 immunoreactivity, but they showed weak or no high‐molecular‐weight MAP2 immunoreactivity. Expression of mRNA containing exons common to high‐ and low‐molecular‐weight MAP2 was detected in the tangentially ellipsoidal cells in the IZ, but expression of mRNA containing an exon specific to high‐molecular‐weight MAP2 was not detected in these cells by in situ hybridization. We interpreted these observations as indicating that the bipolar cells contained MAP2c preferentially, but contained MAP2a and MAP2b (MAP2a/b) at a very low or negligible level. The cells that expressed MAP2c preferentially among the MAP2 splicing variants composed 50% of the preplate cells, most of the MAP2‐positive cells in the hippocampus and the corpus callosum. Double labeling by DiI staining and Dlx2 immunohistochemistry, or by Dlx2 and MAP2 immunohistochemistry, revealed that most of the Dlx2‐positive cells in the IZ expressed MAP2c preferentially at embryonic day 16. Another double‐labeling study revealed that most GAD‐positive cells in the preplate were MAP2a/b positive, whereas most GAD‐positive cells in the IZ expressed MAP2c preferentially, with only a negligible level of MAP2a/b immunoreactivity. We conclude that MAP2 immunoreactivity in the IZ was localized in the tangentially migrating neurons. The tangentially migrating neurons seemed to acquire MAP2a/b immunoreactivity as they entered the preplate or cortical plate and developed into mature neurons. Radially migrating neurons in the IZ were MAP2 negative. After entering to the preplate or the cortical plate, they became MAP2a/b positive as they developed into mature neurons. J. Comp. Neurol. 449:330–342, 2002. © 2002 Wiley‐Liss, Inc.