bluuebunny/crossref_metadata_2025 huggingface dataset:
The value of contrast‐enhanced color doppler ultrasound in the detection of vascularization of finger joints in patients with rheumatoid arthritis
Authors: Klauser Andrea , Frauscher Ferdinand , Schirmer Michael , Halpern Ethan , Pallwein Leo , Herold Manfred , Helweg Gernot , ZurNedden Dieter
Published: October 2002

Objective

A prospective study was performed to assess the usefulness of contrast‐enhanced color Doppler ultrasound (CDUS) in the evaluation of intraarticular vascularization of finger joints in patients with rheumatoid arthritis (RA).

Methods

We investigated 198 finger joints in 46 patients with RA, and 80 finger joints in 10 healthy volunteers. Joints with varying levels of clinical activity of inflammation were classified as being active, moderately active, or inactive. CDUS was performed with a high‐frequency multi‐D linear array transducer. A microbubble‐based ultrasound (US) contrast agent (Levovist; Schering, Berlin, Germany) was intravenously infused. Doppler findings were rated on the basis of both unenhanced and contrast‐enhanced CDUS images.

Results

Healthy joints showed no intraarticular vascularization on either unenhanced or contrast‐enhanced CDUS. Unenhanced CDUS detected intraarticular vascularization in 7 (8%) of 83 inactive joints, in 31 (52%) of 60 moderately active joints, and in 32 (58%) of 55 active joints. Contrast‐enhanced CDUS detected intraarticular vascularization in 41 (49%) of 83 joints with inactive RA, in 59 (98%) of 60 joints with moderately active RA, and in all 55 joints with active RA. Detection of intraarticular vascularization was improved by administration of the microbubble‐based US contrast agent (P < 0.001). Contrast‐enhanced CDUS demonstrated differences in intraarticular vascularization between joints with inactive RA and those with active RA (P < 0.001), between joints with inactive RA and those with moderately active RA (P < 0.001), and between joints with moderately active RA and those with active RA (P < 0.001).

Conclusion

The use of a microbubble‐based US contrast agent significantly improved the detection of intraarticular vascularization in the finger joints of patients with RA. This technique seems to be a useful adjunct in the assessment of disease activity.
Smoking and disease severity in rheumatoid arthritis: Association with polymorphism at the glutathione S‐transferase M1 locus
Authors: Mattey Derek L. , Hutchinson David , Dawes Peter T. , Nixon Nicola B. , Clarke Sheila , Fisher June , Brownfield Ann , Alldersea Julie , Fryer Anthony A. , Strange Richard C.
Published: October 2002

Objective

To determine whether the relationship between smoking and disease severity in women with rheumatoid arthritis (RA) is associated with polymorphism at the glutathione S‐transferase (GST) M1 locus.

Methods

Genotyping for GSTM1 was carried out using polymerase chain reaction methodology on 164 women with established RA. Smoking history was obtained on each patient. Radiographic damage was measured by the Larsen score, and functional outcome was assessed by the Health Assessment Questionnaire (HAQ). Data were analyzed by multiple regression analyses, with correction for age and disease duration.

Results

Ever having smoked was associated with a worse radiographic and functional outcome than was never having smoked. Both past and current smoking were associated with increased disease severity. Stratification by GSTM1 status revealed that polymorphism at this locus affected the relationship between smoking and disease outcome measures. Patients who lacked the GSTM1 gene and had ever smoked had significantly higher Larsen and HAQ scores than did those who lacked the gene and had never smoked. Radiographic outcome in these patients was worse than that in patients who had the GSTM1 gene and who had smoked. The associations were not affected by correction for socioeconomic status. Rheumatoid factor (RF) production was found to be associated with smoking in only the GSTM1‐null patients.

Conclusion

Our data suggest that disease outcome in female RA patients with a history of smoking is significantly worse than in those who have never smoked. Smoking was associated with the most severe disease in patients who carried the GSTM1‐null polymorphism. This association may be due in part to a relationship between the GSTM1 polymorphism and RF production in smokers.
Impaired growth hormone secretion in fibromyalgia patients: Evidence for augmented hypothalamic somatostatin tone
Authors: Paiva Eduardo S. , Deodhar Atul , Jones Kim D. , Bennett Robert
Published: October 2002

Objective

To determine whether female fibromyalgia (FM) patients exhibit a normal growth hormone (GH) response to an acute exercise stressor, and to assess the importance of somatostatin tone in the generation of this GH response.

Methods

Twenty female FM patients were compared with 10 healthy female controls. All subjects exercised to volitional exhaustion on a treadmill. A standard metabolic cart was used to monitor pulse, blood pressure, electrocardiography, oxygen uptake, carbon dioxide output, anaerobic threshold, and maximal workload. Blood was drawn for GH and cortisol measurements 1 hour before exercise, immediately before exercise, immediately after exercise, and 1 hour after exercise. One month later, testing that was exactly similar was performed, except all subjects were given pyridostigmine bromide (Mestinon; 30 mg orally) 1 hour before exercise.

Results

Compared with controls, FM patients failed to exhibit a GH or cortisol response to acute exercise (P = 0.003). After administration of pyridostigmine, 1 hour before exercise, the GH levels of FM patients increased 8‐fold (P = 0.001), to a value comparable with that of controls. Pyridostigmine did not increase the cortisol response to exercise in FM patients. Pyridostigmine alone did not stimulate GH secretion in FM patients, nor did it improve exercise‐induced GH secretion in controls. FM patients with normal insulin‐like growth factor 1 (IGF‐1) levels had an impaired GH response to exercise.

Conclusion

Three new findings are reported: 1) FM patients have a reduced GH response to exercise, 2) pyridostigmine reverses this impaired response, and 3) defective GH secretion in FM can occur in patients with normal IGF‐1 levels. Because pyridostigmine is known to reduce somatostatin tone, it is surmised that the defective GH response to exercise in FM patients probably results from increased levels of somatostatin, a hypothalamic hormone that inhibits GH secretion.
Differential role of neutrophil Fcγ receptor IIIB (CD16) in phagocytosis, bacterial killing, and responses to immune complexes
Authors: Fossati Gianluca , Moots Robert J. , Bucknall Roger C. , Edwards Steven W.
Published: October 2002

Objective

To determine the roles played by the neutrophil Fcγ receptor type II (FcγRII) (CD32) and FcγRIIIb (CD16) in phagocytosis, bacterial killing, and activation by immune complexes (ICs) and to test the hypothesis that inhibition of pathologic effector neutrophil function is possible without compromising host defense.

Methods

Receptor function was probed by enzymic removal of FcγRIIIb from the cell surface and by use of Fab/F(ab‘)2 fragments of monoclonal antibodies to block receptor‐ligand binding. Cells were challenged with (a) serum‐opsonized Staphylococcus aureus, (b) serum‐ and IgG‐opsonized latex particles, and (c) synthetic soluble and insoluble ICs to mimic bacterial and inflammatory stimuli.

Results

Phosphatidylinositol‐phospholipase C treatment removed >97% of surface FcγRIIIb from neutrophils previously treated with tumor necrosis factor α to mobilize intracellular stores of receptor. This treatment profoundly inhibited activation of primed neutrophils by soluble ICs of the type found in diseased rheumatoid joints, but had no effect on phagocytosis and killing of serum‐opsonized S aureus.

Conclusion

FcγRIIIb plays a major role in the secretion of toxic products in response to ICs, but little or no role in the phagocytosis and killing of serum‐opsonized bacteria. The selective suppression of effector neutrophil function is therefore possible. FcγRIIIb, or its intracellular signaling pathway, is a potential therapeutic target in inflammatory diseases such as rheumatoid arthritis, because disruption of its function should decrease inflammatory tissue damage, but not jeopardize host protection against infection.
Anti‐La/SSB antibodies transported across the placenta bind apoptotic cells in fetal organs targeted in neonatal lupus
Authors: Tran Hai B. , Macardle Peter J. , Hiscock Jenny , Cavill Dana , Bradley John , Buyon Jill P. , Gordon Tom P.
Published: October 2002

Objective

To determine whether La and/or Ro epitopes on apoptotic cells in fetal organs that are targeted in neonatal lupus syndrome (NLS) are accessible for binding by autoantibodies in vivo, we traced the fate of transplacental autoantibodies in a murine passive transfer model.

Methods

Pregnant mice at day 15 of gestation (E15) were injected intraperitoneally with human anti‐Ro/La–positive sera or control sera, and transplacental transfer of human autoantibodies was tested by enzyme‐linked immunosorbent assay with recombinant antigens. Multiple cryostat sections at the level of the heart of E17 fetuses were visualized simultaneously for human IgG binding and apoptosis (TUNEL) under confocal microscopy. Serial paraffin sections of E17 and E19 fetuses were examined for histologic evidence of inflammation.

Results

Human IgG anti–52‐kd Ro, anti–60‐kd Ro, and anti‐La autoantibodies were transported efficiently into the fetal circulation. Human IgG–apoptotic cell complexes were detected in the heart (atrial trabeculae and atrioventricular node), skin, liver, and newly forming bone of fetuses from mothers injected with anti‐Ro/La sera but not control sera. The IgG binding was fetal‐specific and organ‐specific; transplacental autoantibodies did not bind to apoptotic cells in the fetal thymus, lung, brain, or gut. The complexes were not associated with an inflammatory reaction. Injection of mothers with affinity‐purified anti‐La autoantibodies (but not anti‐Ro/La Ig depleted of anti‐La) revealed an identical location of IgG binding to apoptotic cells in the fetuses.

Conclusion

This is the first study to demonstrate that transplacental anti‐La autoantibodies bind specifically to apoptotic cells in selected fetal organs in vivo, similar to the organ involvement in NLS. We hypothesize that additional factors are required to promote proinflammatory clearance of IgG–apoptotic cell complexes and subsequent tissue damage.
Regional quantification of cartilage type II collagen and aggrecan messenger RNA in joints with early experimental osteoarthritis
Authors: Matyas John Robert , Huang Dingqiu , Chung May , Adams Mark E.
Published: October 2002

Objective

Accurate assessment of chondrocyte metabolism is a potentially valuable indicator of cartilage health in osteoarthritis (OA). This study was conducted to explore the anabolic metabolism of chondrocytes early in the natural history of an experimental canine model of OA.

Methods

Competitive reverse transcription–polymerase chain reaction was used to calculate the copy number of type II collagen and the messenger RNA (mRNA) levels of aggrecan core protein in articular cartilage samples obtained from different regions of the femorotibial joint 12 and 39 weeks after cruciate transection.

Results

Gene expression of both type II collagen and aggrecan in cartilage samples obtained from experimental joints at both intervals after surgery was elevated significantly compared with that in samples from contralateral control joints. The number of mRNA copies per microgram of DNA of aggrecan exceeded that of type II collagen in control cartilage, but the copy number of type II collagen mRNA exceeded that of aggrecan in OA cartilage. Thus, the ratio of type II collagen–to‐aggrecan mRNA copy number (normalized to DNA) was shown to be characteristically altered in cartilage with experimental OA.

Conclusion

Accurate assessment of multiple gene products in small samples of cartilage taken from focal areas of a joint can be used diagnostically for monitoring chondrocyte metabolism and possibly for staging at least the early phases of this joint disease.
Knee cartilage of spinal cord–injured patients displays progressive thinning in the absence of normal joint loading and movement
Authors: Vanwanseele B. , Eckstein F. , Knecht H. , Stüssi E. , Spaepen A.
Published: October 2002

Objective

Alterations in the morphologic, biochemical, and mechanical properties of cartilage occur after unloading and immobilization in animals. However, the findings have been inconsistent and it is unclear whether such changes also take place in humans. This study tested the hypothesis that progressive thinning of knee joint cartilage is observed after spinal cord injury.

Methods

In this in vivo study, knee cartilage was assessed in patients with complete, traumatic spinal cord injury at 6 (n = 9), 12 (n = 11), and 24 months (n = 6) after injury. Morphologic parameters of the knee cartilage (mean and maximum thickness as well as surface area) were computed from magnetic resonance imaging (MRI) data, and results were compared with those in young, healthy volunteers (n = 9).

Results

After 6 months of injury, the mean articular‐cartilage thickness was significantly less in the patella and medial tibia (decrease of 10% and 16%, respectively; P < 0.05), but not in the lateral tibia (decrease of 10%), compared with the MRI findings in healthy volunteers. After 12 and 24 months of injury, the differences amounted to a reduction of 21% and 23%, respectively, in the patella, 24% and 25%, respectively, in the medial tibia, and 16% and 19%, respectively, in the lateral tibia. The changes were significant in all 3 surfaces of the spinal cord–injured joint cartilage (P < 0.05–0.01).

Conclusion

Our data show, for the first time, that progressive thinning (atrophy) of human cartilage occurs in the absence of normal joint loading and movement. This may have important implications for patient management, in particular for spinal cord–injured patients and patients who are immobilized after surgery.
Preferences for improved health examined in 1,024 patients with rheumatoid arthritis: Pain has highest priority
Authors: Heiberg Turid , Kvien Tore K.
Published: October 2002

Objective

To examine preferences for improved health in patients with rheumatoid arthritis (RA).

Methods

A survey was mailed to patients with RA enrolled in a county‐based register. The questionnaire comprised a variety of health status measures (Medical Outcome Study Short Form‐36, Arthritis Impact Measurement Scales 2 [AIMS2], Modified Health Assessment Questionnaire, and visual analog scale for pain and fatigue). The patients were asked to check 3 of 12 areas in which they would most like to see improvement (item 60 AIMS2). The number of respondents was 1,024 (mean age/disease duration 63.4/12.7 years, 78.7% female).

Results

Pain was the preferred area for improvement in all subgroups of patients. Preference for improvement in pain was associated with lower age, higher levels of perceived pain, and lower scores for self efficacy related to pain. One‐third of the patients with this preference did not report use of pain‐relieving medication.

Conclusion

Pain is the area of health in which almost 70% of the patients would like to see improvement. This study suggests that more attention should be paid to the examination of patient preferences for improvement in health.
Comparison of synovial tissues from the knee joints and the small joints of rheumatoid arthritis patients: Implications for pathogenesis and evaluation of treatment
Authors: Kraan Maarten C. , Reece Richard J. , Smeets Tom J. M. , Veale Douglas J. , Emery Paul , Tak Paul P.
Published: October 2002

Objective

Serial synovial biopsy samples are increasingly being used for the evaluation of novel therapies for rheumatoid arthritis (RA). Most studies have used tissues from knee biopsies, but technical improvements have made serial small joint arthroscopy feasible as well. Theoretically, there could be differences in the features of synovial inflammation between various joints as a result of mechanical factors, differences in innervation, and other factors. We therefore undertook this study to compare the cell infiltrate in paired synovial biopsy samples from inflamed knee joints and paired inflamed small joints of patients with RA.

Methods

Nine RA patients with both an inflamed knee joint and an inflamed small joint (wrist or metacarpophalangeal joint) underwent an arthroscopic synovial biopsy of both joints on the same day. Multiple biopsy specimens were collected and stained for macrophages, T cells, plasma cells, fibroblast‐like synoviocytes, and interleukin‐6 (IL‐6) by immunohistochemistry. Sections were evaluated by digital image analysis.

Results

There were no significant differences in mean cell numbers for all markers investigated in samples from the knee joint compared with samples from the small joints. We detected statistically significant correlations for the numbers of sublining macrophages, T cells, and plasma cells, as well as for IL‐6 expression, between the knee joint and the small joints. However, there was no significant correlation between different joints for the numbers of intimal macrophages or fibroblast‐like synoviocytes.

Conclusion

The results of this study show that the inflammation in one inflamed joint is generally representative of that in other inflamed joints. Therefore, it is possible to use serial samples from the same joint, selecting either large or small joints, for the evaluation of antirheumatic therapies.
Information seeking and mediated searching: Part 2. Uncertainty and its correlates
Authors: Wilson T.D. , Ford N.J. , Ellis D. , Foster A.E. , Spink A.
Published: October 2002

This article explores the relationship between the concept of uncertainty in information seeking, within a model of the problem‐solving process proposed by Wilson (1999a) and variables derived from other models and from the work of Ellis and Kuhlthau. The research has involved longitudinal data collection in the United States and United Kingdom employing three interview schedules (incorporating self‐completed questionnaires) used for pre‐ and postsearch interviews: and postsearch interviews with the information seeker and the search intermediary. In addition, the Sheffield team employed a fourth set of instruments in a follow‐up interview some 2 months after the search. Related search episodes, with a professional search intermediary using the Dialog Information Service and other sources were audiotaped, and search transaction logs were recorded. The mediated search clients were faculty and research students engaged in either personal or externally supported research projects. The article concludes that the problem solving model is recognized by such researchers as describing their activities and that the uncertainty concept, operationalized as here, serves as a useful variable in understanding information‐seeking behavior. It also concludes that Ellis's concept of “search characteristics” and Kuhlthau's information‐seeking stages are independent of the problem stage, and that a set of affective variables, based on those of Kuhlthau, appear to signify a generalized positive or negative affective orientation towards the course of the information problem solution.
Information seeking and mediated searching. Part 4. Cognitive styles in information seeking
Authors: Ford Nigel , Wilson T.D. , Foster Allen , Ellis David , Spink Amanda
Published: October 2002

This is the fourth in a series resulting from a joint research project directed by Professor Tom Wilson in the United Kingdom and Dr. Amanda Spink in the United States. The analysis reported here sought to test a number of hypotheses linking global/analytic cognitive styles and aspects of researchers' problem‐solving and related information‐seeking behavior. One hundred and eleven postdoctoral researchers were assessed for Witkin's field dependence/independence using Riding's Cognitive Styles Analysis and for Pask's holist/serialist biases using items from Ford's Study Processes Questionnaire. These measures were correlated with the researchers' perceptions of aspects of their problem‐solving and information‐seeking behavior, and with those of the search intermediary who performed literature searches on their behalf. A number of statistically significant correlations were found. Field‐independent researchers were more analytic and active than their field‐dependent counterparts. Holists engaged more in exploratory and serendipitous behavior, and were more idiosyncratic in their communication than serialists.
Information interaction: Providing a framework for information architecture
Authors: Toms Elaine G.
Published: October 2002

Information interaction is the process that people use in interacting with the content of an information system. Information architecture is a blueprint and navigational aid to the content of information‐rich systems. As such information architecture performs an important supporting role in information interactivity. This article elaborates on a model of information interactivity that crosses the “no‐man's land” between user and computer articulating a model that includes user, content and system, illustrating the context for information architecture.
A case study of information‐seeking behavior in 7‐year‐old children in a semistructured situation
Authors: Cooper Linda Z.
Published: October 2002

This article presents a case study of the information‐seeking behavior of 7‐year‐old children in a semistructured situation in their school library media center. The study focuses on how young children who are in the process of learning to read cope with searching for information in a largely textual corpus, and how they make up for their deficit in textual experience. Children's search strategies are examined and discussed in the context of computer versus shelf searching, textual versus visual searching, and in comparison with adult search dimensions previously established.
Mustering motivation to enact decisions: how decision process characteristics influence goal realization
Authors: Dholakia Utpal M. , Bagozzi Richard P.
Published: October 2002

Decision scientists tend to focus mainly on decision antecedents, studying how people make decisions. Action psychologists, in contrast, study post‐decision issues, investigating how decisions, once formed, are maintained, protected, and enacted. Through the research presented here, we seek to bridge these two disciplines, proposing that the process by which decisions are reached motivates subsequent pursuit and benefits eventual realization. We identify three characteristics of the decision process (DP) as having motivation‐mustering potential: DP effort investment, DP importance, and DP confidence. Through two field studies tracking participants' decision processes, pursuit and realization, we find that after controlling for the influence of the motivational mechanisms of goal intention and implementation intention, the three decision process characteristics significantly influence the successful enactment of the chosen decision directly. The the oretical and practical implications of these findings are considered and future research opportunities are identified. Copyright © 2002 John Wiley & Sons, Ltd.
The role of calcium‐binding proteins in the control of transcription: structure to function
Authors: Ikura Mitsuhiko , Osawa Masanori , Ames James B.
Published: October 2002

Transcriptional regulation is coupled with numerous intracellular signaling processes often mediated by second messengers. Now, growing evidence points to the importance of Ca2+, one of the most versatile second messengers, in activating or inhibiting gene transcription through actions frequently mediated by members of the EF‐hand superfamily of Ca2+‐binding proteins. Calmodulin and calcineurin, representative members of this EF‐hand superfamily, indirectly regulate transcription through phosphorylation/dephosphorylation of transcription factors in response to a Ca2+ increase in the cell. Recently, a novel EF‐hand Ca2+‐binding protein called DREAM has been found to interact with regulatory sequences of DNA, thereby acting as a direct regulator of transcription. Finally, S100B, a dimeric EF‐hand Ca2+‐binding protein, interacts with the tumor suppressor p53 and controls its transcriptional activity. In light of the structural studies reported to date, this review provides an overview of the structural basis of EF‐hand Ca2+‐binding proteins linked with transcriptional regulation. BioEssays 24:625–636, 2002. © 2002 Wiley Periodicals, Inc.
Conformational control through translocational regulation: a new view of secretory and membrane protein folding
Authors: Lingappa Vishwanath R. , Rutkowski D. Thomas , Hegde Ramanujan S. , Andersen Olaf S.
Published: October 2002

We suggest a new view of secretory and membrane protein folding that emphasizes the role of pathways of biogenesis in generating functional and conformational heterogeneity. In this view, heterogeneity results from action of accessory factors either directly binding specific sequences of the nascent chain, or indirectly, changing the environment in which a particular domain is synthesized. Entrained by signaling pathways, these variables create a combinatorial set of necessary‐but‐not‐sufficient conditions that enhance synthesis and folding of particular alternate, functional, conformational forms. We therefore propose that protein conformation is productively regulated by the cell during translocation across the endoplasmic reticulum (ER), a concept that may account for currently poorly understood aspects of physiological function, natural selection, and disease pathogenesis. BioEssays 24:741–748, 2002. © 2002 Wiley Periodicals, Inc.
Mitochondrial dysfunction and Down's syndrome
Authors: Arbuzova Svetlana , Hutchin Tim , Cuckle Howard
Published: October 2002

Neither the pathogenesis nor the aetiology of Down's syndrome (DS) are clearly understood. Numerous studies have examined whether clinical features of DS are a consequence of specific chromosome 21 segments being triplicated. There is no evidence, however, that individual loci are responsible, or that the oxidative damage in DS could be solely explained by a gene dosage effect. Using astrocytes and neuronal cultures from DS fetuses, a recent paper shows that altered metabolism of the amyloid precursor protein and oxidative stress result from mitochondrial dysfunction.1 These findings are consistent with considerable data implicating the role of the mitochondrial genome in DS pathogenesis and aetiology. BioEssays 24:681–684, 2002. © 2002 Wiley Periodicals, Inc.
Surface enhanced IR absorption of nucleic acids from tumor cells: FTIR reflectance study
Authors: Dovbeshko G. I. , Chegel V. I. , Gridina N. Y. , Repnytska O. P. , Shirshov Y. M. , Tryndiak V. P. , Todor I. M. , Solyanik G. I.
Published: October 2002

The data on surface enhanced IR absorption (SEIRA) of nucleic acids deposited on a metal substrate were obtained using FTIR in reflectance mode. A 200–400 Å thick gold film on a glass plate was the metal substrate. The approximate enhancement factors of the SEIRA for different vibrations of nucleic acids in our experimental conditions were 3–5. The roughness of the Au surface was about 50 Å. Application of this method to nucleic acids isolated from tumor cells revealed some possible peculiarities of their structural organization, namely, the appearance of unusual sugar and base conformations, modification of the phosphate backbone, redistribution of the H‐bond net, and so forth. This method enhanced a set of the bands, which is impossible to observe in conventional IR geometry. The SEIRA spectra of the RNA from tumor cells showed more sensitivity to the grade of tumor malignancy than the spectra of the DNA. After application of the anticancer drug doxorubicin to sensitive and resistant strains, the DNA isolated from these strains had different spectral features, especially in the region of the phosphate I and II bands. As induced by anticancer drugs, the conformational changes in the DNA from resistant and sensitive cancer strains could be characterized with different levels of structure disordering. © 2002 Wiley Periodicals, Inc. Biopolymers (Biospectroscopy) 67: 470–486, 2002
Effects of terbium and pH on structure of anticoagulation factor II from Agkistrodon acutus venom probed by fluorescent spectroscopy and equilibrium dialysis
Authors: Xu Xiao Long , Liu Qing Liang , Wu Bin , Xie Yong Shu
Published: October 2002

Anticoagulation factor II (ACF II) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X‐binding protein with marked anticoagulant activity. Present studies show that the pH has a marked effect on the fluorescence intensity of holo‐ACF II; however, no appreciable shift of the emission maximum of holo‐ACF II was observed in the pH range of 3–10. It was deduced from a relatively weak fluorescence emission of holo‐ACF II at a neutral pH (6–7) that native holo‐ACF II assumes a compactly folded structure in which the most interior Trp residues and quenchers are adjacent. Terbium ions can completely replace both Ca2+ ions in holo‐ACF II as determined by equilibrium dialysis. Two Tb3+‐binding sites with different apparent Tb3+ association constant values, (2.1 ± 0.2) and (1.0 ± 0.1) × 107
M−1, were identified through Tb3+ fluorescence titration. In addition, it was confirmed from the titration of holo‐ACF II and Tb3+‐ACF II with N‐bromosuccinimide (NBS) that only interior Trp residues are involved in the energy transfer to Tb3+ ions and all accessible Trp residues located in the surface of holo‐ACF II have a similar affinity to NBS while those located in the surface of Tb3+‐ACF II have two different kinds of affinity to NBS, which suggests a conformational change of holo‐ACF II on the substitution of Tb3+ for Ca2+. © 2002 Wiley Periodicals, Inc. Biopolymers (Biospectroscopy) 67: 387–393, 2002
Metabolic pathway analysis of a recombinant yeast for rational strain development
Authors: Carlson Ross , Fell David , Srienc Friedrich
Published: October 2002

Elementary mode analysis has been used to study a metabolic pathway model of a recombinant Saccharomyces cerevisiae system that was genetically engineered to produce the bacterial storage compound poly‐β‐hydroxybutyrate (PHB). The model includes biochemical reactions from the intermediary metabolism and takes into account cellular compartmentalization as well as the reversibility/irreversibility of the reactions. The reaction network connects the production and/or consumption of eight external metabolites including glucose, acetate, glycerol, ethanol, PHB, CO2, succinate, and adenosine triphosphate (ATP). Elementary mode analysis of the wild‐type S. cerevisiae system reveals 241 unique reaction combinations that balance the eight external metabolites. When the recombinant PHB pathway is included, and when the reaction model is altered to simulate the experimental conditions when PHB accumulates, the analysis reveals 20 unique elementary modes. Of these 20 modes, 7 produce PHB with the optimal mode having a theoretical PHB carbon yield of 0.67. Elementary mode analysis was also used to analyze the possible effects of biochemical network modifications and altered culturing conditions. When the natively absent ATP citrate‐lyase activity is added to the recombinant reaction network, the number of unique modes increases from 20 to 496, with 314 of these modes producing PHB. With this topological modification, the maximum theoretical PHB carbon yield increases from 0.67 to 0.83. Adding a transhydrogenase reaction to the model also improves the theoretical conversion of substrate into PHB. The recombinant system with the transhydrogenase reaction but without the ATP citrate‐lyase reaction has an increase in PHB carbon yield from 0.67 to 0.71. When the model includes both the ATP citrate‐lyase reaction and the transhydrogenase reaction, the maximum theoretical carbon yield increases to 0.84. The reaction model was also used to explore the possibility of producing PHB under anaerobic conditions. In the absence of oxygen, the recombinant reaction network possesses two elementary modes capable of producing PHB. Interestingly, both modes also produce ethanol. Elementary mode analysis provides a means of deconstructing complex metabolic networks into their basic functional units. This information can be used for analyzing existing pathways and for the rational design of further modifications that could improve the system's conversion of substrate into product. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 121–134, 2002.
Ultra scaledown to predict filtering centrifugation of secreted antibody fragments from fungal broth
Authors: Boulding N. , Yim S. S. S. , Keshavarz‐Moore E. , Ayazi Shamlou P. , Berry M.
Published: October 2002

Extracellularly expressed anti–hen egg lysozyme single‐chain antibody fragments (scFv) produced by Aspergillus awamori were recovered using filtering centrifugation. Two filtering centrifuges with 0.5‐ and 30‐L capacities were used to represent laboratory‐ and pilot‐scale equipment, respectively. Critical regime analysis using the computational fluid dynamics (CFD) technique provided information about the local energy dissipation rates in both units. Experimental data indicated loss of scFv activity for energy dissipation rates above about 2.0 × 104 W kg−1. This loss of activity increased in the presence of gas–liquid interfaces during filtering centrifugation. An ultra scaledown filtering centrifuge with a maximum working volume of 35 mL was designed to mimic the operating conditions identified by the critical regime analysis for the laboratory‐ and pilot‐plant‐scale units. The recovered scFv activity levels and the separation performance of the three units were comparable when operated at equal maximum energy dissipation rates. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 381–388, 2002.
Optimal temperature and photoperiod for the cultivation of Agardhiella subulata microplantlets in a bubble‐column photobioreactor
Authors: Huang Yao‐ming , Rorrer Gregory L.
Published: October 2002

The optimal temperature and illumination photoperiod requirements for the phototrophic growth of a novel microplantlet suspension culture derived from the macrophytic marine red alga Agardhiella subulata were determined. The optimal growth temperature was 24°C. The effects of illumination light‐dark (LD) photoperiod (hour of light:hours of darkness within a 24 h cycle) on biomass production was studied within a bubble‐column photobioreactor. The 4.5 cm diameter photobioreactor was maintained at near‐saturation conditions with respect to light flux (38 μmol photons m−2 s−1), nutrient medium delivery (20% nutrient replacement per day), and CO2 delivery (0.35 mmol CO2 L−1 h−1) so that the cumulative effects of photodamage on the cell density versus time curve at photoperiods approaching continuous light could be observed. Biomass production was maximized at 16:8 LD, where biomass densities exceeding 3.6 g dry cell mass L−1 were achieved after 60 days in culture. Biomass production was proportional to photoperiod at low fractional photoperiods (≤10:14 LD), but high fractional photoperiods approaching continuous light (≥ 20:4 LD) shut down biomass production. Biomass production versus time profiles under resource‐saturated cultivation conditions were adequately described by a cumulative photodamage growth model, which coupled reversible photodamage processes to the specific growth rate. Under light‐saturated growth conditions, the rate constant for photodamage was kd = 1.17 ± 0.28 day−1 (±1.0 SE), and the rate constant for photodamage repair was kr = 5.12 ± 0.95 day−1 (±1.0 SE) at 24°C. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 135–144, 2002.
Biodegradation of crude oil across a wide range of salinities by an extremely halotolerant bacterial consortium MPD‐M, immobilized onto polypropylene fibers
Authors: Díaz María Piedad , Boyd Kenneth G. , Grigson Steve J. W. , Burgess J. Grant
Published: October 2002

The bacterial consortium MPD‐M, isolated from sediment associated with Colombian mangrove roots, was effective in the treatment of hydrocarbons in water with salinities varying from 0 to 180 g L−1. Where the salinity of the culture medium surpassed 20 g L−1, its effectiveness increased when the cells were immobilized on polypropylene fibers. Over the range of salinity evaluated, the immobilized cells significantly enhanced the biodegradation rate of crude oil compared with free‐living cells, especially with increasing salinity in the culture medium. Contrary to that observed in free cell systems, the bacterial consortium MPD‐M was highly stable in immobilized systems and it was not greatly affected by increments in salinity. Biodegradation was evident even at the highest salinity evaluated (180 g L−1), where biodegradation was between 4 and 7 times higher with immobilized cells compared to free cells. The biodegradation of pristane (PR) and phytane (PH) and of the aromatic fraction was also increased using cells immobilized on polypropylene fibers. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 145–153, 2002.
The measurement of Bacillus mycoides spore adhesion using atomic force microscopy, simple counting methods, and a spinning disk technique
Authors: Bowen W. Richard , Fenton Adam S. , Lovitt Robert W. , Wright Chris J.
Published: October 2002

An atomic force microscope has been used to study the adhesion of Bacillus mycoides spores to a hydrophilic glass surface and a hydrophobic‐coated glass surface. AFM images of spores attached to the hydrophobic‐coated mica surface allowed the measurement of spore dimensions in an aqueous environment without desiccation. The spore exosporium was observed to be flexible and to promote the adhesion of the spore by increasing the area of spore contact with the surface. Results from counting procedures using light microscopy matched the density of spores observed on the hydrophobic‐coated glass surface with AFM. However, no spores were observed on the hydrophilic glass surface with AFM, a consequence of the weaker adhesion of the spores at this surface. AFM was also used to quantify directly the interactions of B. mycoides spores at the two surfaces in an aqueous environment. The measurements used “spore probes” constructed by immobilizing a single spore at the apex of a tipless AFM cantilever. The data showed that stretching and sequential bond breaking occurred as the spores were retracted from the hydrophilic glass surface. The greatest spore adhesion was measured at the hydrophobic‐coated glass surface. An attractive force on the spores was measured as the spores approached the hydrophobic‐coated surface. At the hydrophilic glass surface, only repulsive forces were measured during the approach of the spores. The AFM force measurements were in qualitative agreement with the results of a hydrodynamic shear adhesion assay that used a spinning disk technique. Quantitatively, AFM measurements of adhesive force were up to 4 × 103 times larger than the estimates made using the spinning disk data. This is a consequence of the different types of forces applied to the spore in the different adhesion assays. AFM has provided some unique insights into the interactions of spores with surfaces. No other instrument can make such direct measurements for single microbiological cells. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 170–179, 2002
Advantages of continuous over batch reactors for the kinetic analysis of enzymes inhibited by an unknown substrate impurity
Authors: Gallifuoco Alberto , Alfani Francesco , Cantarella Maria
Published: October 2002

A new experimental technique, employing a continuous stirred‐tank reactor, for studying enzyme kinetics in the presence of inhibitor‐contaminated substrate is described. The proposed method is simulated mathematically for competitive, uncompetitive, and mixed‐type noncompetitive inhibition. The step‐by‐step experimental procedure is described, as is the necessary data analysis for determining the kinetic parameters. Differences in system response for enzyme inhibition by excess substrate and by an impurity are illustrated, and a stability analysis of the system is performed. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 641–646, 2002.
High‐activity enzyme‐polyurethane coatings
Authors: Drevon Géraldine F. , Danielmeier Karsten , Federspiel William , Stolz Donna B. , Wicks Douglas A. , Yu Poli C. , Russell Alan J.
Published: October 2002

The synthesis of water‐borne polyurethane coatings in the presence of diisopropylfluorophosphatase (DFPase, E.C. 3.8.2.1) enabled the irreversible attachment of the enzyme to the polymeric matrix. The distribution of immobilized DFPase as well as activity retention are homogeneous within the coating. The resulting enzyme‐containing coating (ECC) film hydrolyzes diisopropylfluorophosphate (DFP) in buffered media at high rates, retaining approximately 39% intrinsic activity. Decreasing ECC hydrophilicity, via the use of a less hydrophilic polyisocyanate during polymerization, significantly enhanced the intrinsic activity of the ECC. DFPase‐ECC has biphasic deactivation kinetics, where the initial rapid deactivation of DFPase‐ECC leads to the formation of a hyperstable and active form of enzyme. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 785–794, 2002.
A functional genomics approach using metabolomics and in silico pathway analysis
Authors: Förster Jochen , Gombert Andreas Karoly , Nielsen Jens
Published: October 2002

In the field of functional genomics increasing effort is being undertaken to analyze the function of orphan genes using metabolome data. Improved analytical equipment allows screening simultaneously for a high number of metabolites. Such metabolite profiles are analyzed using multivariate data analysis techniques and changes in the genotype will in many cases lead to different metabolite profiles. Here, a theoretical framework that may be applied to identify the function of orphan genes is presented. The approach is based on a combination of metabolome analysis combined with in silico pathway analysis. Pathway analysis may be carried out using convex analysis and a change in the active pathway structure of deletion mutants expressed in a different metabolite profile may disclose the function or the functional class of an orphan gene. The concept is illustrated using a simplified model for growth of Saccharomyces cerevisiae. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 703–712, 2002.
Achieving community justice through faith‐based initiatives
Authors: Small Mark A.
Published: October 2002

Prior to 1996, most churches and other faith‐based organizations were ineligible to receive federal funding for community services. In a little noticed provision of the Welfare Reform Act of 1996 known as ‘charitable choice,’ the federal government allowed religious groups to receive money for social programs without requiring them to censor their religious expression or give up their religious identity. States, with varying degrees of vigor and success, have partnered with faith‐based organizations to provide community services that serve the purpose of transitioning people from welfare to work. Recent political developments and legislation suggest an expanded role for faith‐based organizations to receive federal money to develop community services, including money for community justice projects. The Comment begins with an overview of the political and legal contexts that allow the development of promoting community justice initiatives through faith‐based efforts. Next, there is a discussion of practical considerations that influence decisions of churches and faith‐based organizations to participate in these activities. Finally, there is discussion of the roles social scientists and others might play to facilitate community justice initiatives through faith‐based efforts. Copyright © 2002 John Wiley & Sons, Ltd.
General purpose software package for electromagnetics engineering education
Authors: Bértolo J. M. , Obelleiro F. , Taboada J. M. , Rodríguez J. L.
Published: October 2002

This article presents a general‐purpose educational software applied to electromagnetics engineering education. The program presented allows the students to obtain the RCS, current density, and near field distributions of perfectly electric conducting (PEC) canonical geometries being illuminated by arbitrary incident plane waves. © 2002 Wiley Periodicals, Inc. Comput Appl Eng Educ 10: 33–44, 2002; Published online in Wiley InterScience (www.interscience.wiley.com.); DOI 10.1002/cae.10015
Management of a large pseudoaneurysm secondary to balloon dilation for native coarctation of the aorta with coil occlusion after stent implantation in a child
Authors: Pedra Carlos A.C. , Pilla Carlo B. , Braga Sérgio L.N. , Esteves César A. , Fontes Valmir F.
Published: October 2002

We report a case of a 9‐year‐old girl status post–balloon dilation for native coarctation of the aorta who had a large aortic pseudoaneurysm 1 year after the initial procedure. The aneurysm was occluded with multiple coils after stent implantation to the aorta. The technique and possible advantages of this novel approach are discussed. Cathet Cardiovasc Intervent 2002;56:262–266. © 2002 Wiley‐Liss, Inc.
Mechanism of cutting balloon angioplasty for in‐stent restenosis: An intravascular ultrasound study
Authors: Montorsi Piero , Galli Stefano , Fabbiocchi Franco , Loaldi Alessandro , Trabattoni Daniela , Grancini Luca , Cozzi Sergio , Ravagnani Paolo , Parodi Oberdan , Bartorelli Antonio L.
Published: October 2002

We investigated by intravascular ultrasound (IVUS) the mechanism of action of cutting balloon (CB) angioplasty in patients with in‐stent restenosis. Seventy‐one consecutive restenotic lesions of 66 patients were studied by quantitative coronary angiography (QCA) and IVUS before, immediately after, and, in 20 cases, at 24‐hr time interval after CB. CB was selected according to 1:1 CB‐to‐stent ratio and inflated at 8 atm for 60–90 sec. Both IVUS planar and volumetric (Simpson's rule, 25 patients) analysis were carried out. IVUS measurements included external elastic membrane area (EEMA), stent area (SA), minimal lumen area (MLA), and restenosis area (RA). Following CB, QCA analysis showed increase of minimal lumen diameter (1.17 ± 0.46 vs. 2.45 ± 0.51 mm; P < 0.0001) and decrease of diameter stenosis (64% ± 13% vs. 21% ± 9%; P < 0.0001). IVUS measurements showed a significant increase of MLA (2.18 ± 0.80 vs. 7.31 ± 1.8 mm2; P < 0.0001), SA (9.62 ± 2.6 vs. 10.7 ± 2.75 mm2; P < 0.0001), and EEMA (17.27 ± 5 vs. 18.1 ± 5 mm2; P < 0.0001) and a decrease of RA (7.43 ± 2.63 vs. 3.45 ± 1.39 mm2; P < 0.0001). No significant change was observed in the original plaque + media area (7.65 ± 3 vs. 7.38 ± 2.9 mm2; P = NS). Thus, of the total lumen enlargement (5.13 ± 1.85 mm2), 23% was the result of increase in mean SA, whereas 77% was the result of a decrease in mean RA. These changes were associated with a 5% increase in EEMA. IVUS volumetric changes paralleled planar variations. Angiographic and IVUS changes were well maintained at 24 hr. CB enlarges coronary lumen mainly by in‐stent tissue reduction associated with a moderate degree of additional stent expansion. Favorable QCA and IVUS acute results are maintained at 24 hr. Cathet Cardiovasc Intervent 2002;56:166–173. © 2002 Wiley‐Liss, Inc.