bluuebunny/crossref_metadata_2025 huggingface dataset:
Stability of crystalline proteins
Authors: Shenoy Bhami , Wang Yi , Shan Weizhong , Margolin Alexey L.
Published: August 2002

By using two model proteins, glucose oxidase and lipase, we demonstrate that dry crystalline formulations are significantly more stable than their amorphous counterparts. The results of Fourier‐transform infrared spectroscopy indicate that crystalline proteins better maintain their native conformation in accelerated stability studies. The lower tendency of crystalline proteins to aggregate is confirmed by size‐exclusion chromatography. The data suggest that protein crystallization may significantly improve some aspects of protein handling, and change the way biopharmaceuticals are produced, formulated, and delivered. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 73: 358–369, 2001.
Effects of oxygenation and flow on the viability and function of rat hepatocytes cocultured in a microchannel flat‐plate bioreactor
Authors: Tilles Arno W. , Baskaran Harihara , Roy Partha , Yarmush Martin L. , Toner Mehmet
Published: August 2002

The goal of this study was to investigate the viability and synthetic function of rat hepatocytes cocultured with 3T3‐J2 fibroblasts in a small‐scale microchannel flat‐plate bioreactor with and without an internal membrane oxygenator under flow. Bioreactor channel heights ranged between 85 and 500 μm and medium flow rates ranged between 0.06 and 4.18 mL/min. The results showed that the bioreactor without the oxygenator resulted in significantly decreased viability and function of hepatocytes, whereas hepatocytes in the bioreactor with internal membrane oxygenator were able to maintain their viability and function. The shear stress calculations showed that, at lower wall shear stresses (0.01 to 0.33 dyn/cm2), hepatocyte functions, measured as albumin and urea synthesis rates, were as much as 2.6‐ and 1.9‐fold greater, respectively, than those at higher wall shear stresses (5 to 21 dyn/cm2). Stable albumin and urea synthesis rates for 10 days of perfusion were also demonstrated in the bioreactor with internal membrane oxygenator. These results are relevant in the design of hepatocyte bioreactors and the eventual scaling‐up to clinical devices. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 73: 379–389, 2001.
Intra‐particle oxygen diffusion limitation in solid‐state fermentation
Authors: Oostra J. , le Comte E. P. , van den Heuvel J. C. , Tramper J. , Rinzema A.
Published: August 2002

Oxygen limitation in solid‐state fermentation (SSF) has been the topic of modeling studies, but thus far, there has been no experimental elucidation on oxygen‐transfer limitation at the particle level. Therefore, intra‐particle oxygen transfer was experimentally studied in cultures of Rhizopus oligosporus grown on the surface of solid, nutritionally defined, glucose and starch media. The fungal mat consisted of two layers—an upper layer with sparse aerial hyphae and gas‐filled interstitial pores, and a dense bottom layer with liquid‐filled pores. During the course of cultivation ethanol was detected in the medium indicating that oxygen was depleted in part of the fungal mat. Direct measurement of the oxygen concentrations in the fungal mat during cultivation, using oxygen microelectrodes, showed no oxygen depletion in the upper aerial layer, but revealed development of steep oxygen concentration gradients in the wet bottom layer. Initially, the fungal mat was fully oxygenated, but after 36.5 hours oxygen was undetectable at 100 μm below the gas–liquid interface. This was consistent with the calculated oxygen penetration depth using a reaction‐diffusion model. Comparison of the overall oxygen consumption rate from the gas phase to the oxygen flux at the gas‐liquid interface showed that oxygen consumption of the microorganisms occurred mainly in the wet part of the fungal mat. The contribution of the aerial hyphae to overall oxygen consumption was negligible. It can be concluded that optimal oxygen transfer in SSF depends on the available interfacial gas–liquid surface area and the thickness of the wet fungal layer. It is suggested that the moisture content of the matrix affects both parameters and, therefore, plays an important role in optimizing oxygen transfer in SSF cultures. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 13–24, 2001.
In situ product removal of ketoses by immobilized 3‐amino phenyl boronic acid: Effect of immobilization method on pH profile
Authors: Dukler Avinoam , Freeman Amihay
Published: August 2002

The use of polymeric derivatives of phenylboronic acid (PBA) as an effective means for specific in situ product removal of ketoses from aldose‐containing reaction mixtures, strongly depends on the retention of selective binding of ketoses exhibited by soluble PBA and 3‐amino PBA, by their polymeric, water insoluble analogs. In this communication we demonstrate that immobilization chemistry has a strong effect on ketose preferred binding by polymeric PBA derivatives. Our results indicate that for the preparation of an effective and more specific adsorbent, 3‐amino PBA should be coupled to the polymeric carrier via alkylamino chemistry and not via the commonly employed amido derivative. Immobilized alkylamino‐PBA exhibited selective fructose and xylulose binding throughout glucose and xylose isomerization processes at the pH range of 7.0–8.0. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 25–28, 2001.
Analysis of menthol in three traditional Chinese medicinal herbs and their compound formulation by GC‐MS
Authors: Lin Ruimin , Tian Jing , Huang Ge , Li Tiechun , Li Famei
Published: August 2002

A GC‐MS method was developed for identification and determination of menthol in three traditional Chinese medicinal herbs and their compound formulation, a granule for treating colds. Volatile oil was simultaneously distilled and extracted into ethyl ether in a unique glass extractor. The separation was performed on an HP‐5 MS column. The standard addition method was used for quantitative determination of menthol content in herbal materials and in the granule. A component in the samples was chosen as the internal standard. The contents were calculated with the ratio of peak area percentage. Menthol was identified as the main component in Mentha haplocalyx Briq., and also existed in spikes of Schizonepeta tenuifolia Briq., Folium perilla frutescens (L.) Britt. and granules. The quantitative calibration range was 0.21–10.5 mg/mL. Good precision was demonstrated by an RSD < 4.0%. The mean content of menthol in Mentha haplocalyx Briq., spikes of Schizonepeta tenuifolia Briq., Folium perilla frutescens (L.) Britt. and granules was 121, 0.234, 1.03 and 1.84 mg/kg respectively. Copyright © 2002 John Wiley & Sons, Ltd.
Conceptualizing and studying the contribution of networks in environmental management and sustainable development
Authors: Roome Nigel
Published: August 2002

This special issue ofBusiness Strategy and the Environmentexamines the role and contribution of networks to environmental management and sustainable development. The paper has two main purposes. It situates networks and their contribution to environmental management and sustainable development within the tradition of meta‐problems and socio‐technical systems. This provides an important conceptual frame for the field. It offers a robust setting within which a number of other theories can be set. The need to establish this conceptual framing is particularly important, as the interest in networks from researchers and practitioners has progressed rapidly during the past five years. Moreover, it is anticipated that this interest in networks will continue to develop as the focus of attention moves from issues of environmental management, to sustainable development, and then on to the association between sustainable development and other forms of global change. The significance of these and other trends is identified. The paper also introduces the other four contributions to this special issue, which examine the role of networks from a number of theoretical and empirical perspectives. The paper concludes by drawing out a set of research themes that appear to deserve critical attention in the future. Copyright © 2001 John Wiley & Sons, Ltd. and ERP Environment
Employment impacts of cleaner production – evidence from a German study using case studies and surveys
Authors: Pfeiffer Friedhelm , Rennings Klaus
Published: August 2002

The study assesses net employment effects of technical progress, which can be expected by the ongoing transition from end‐of‐pipe technologies towards cleaner production. Empirical evidence is presented on the basis of case studies and firm data including a telephone survey from German industry. The main result is that the transition from end‐of‐pipe technologies to cleaner production leads to a net creation of jobs, which is however restricted to a only small number of firms and to the group of highly skilled labour. Eco‐innovations, like other innovations, are non‐neutral. The demand for skilled and highly skilled labour rises while the demand for unskilled labour decreases. Synergies between environmental, labour market and innovation policy are apparent but they are however small and specific. The exploitation of these synergies requires the design of specific policy programmes differentiating between types of eco‐innovation. The promotion of product‐integrated environmental measures should be more successful if new products complement older ones, while process‐integrated environmental measures should be more successful if consumers' demand is more price elastic. Copyright © 2001 John Wiley & Sons, Ltd. and ERP Environment
What's in it for the customers? Successfully marketing green clothes
Authors: Meyer Arnt
Published: August 2002

Most green marketing literature is strongly based on the assumption that a consumer's environmental awareness is a pre‐condition for green purchasing. While following such a ‘behaviouristic’ green marketing both scholars and companies seem to have neglected that green products are bought only if customers perceive the products as superior to competitors’ offerings. This paper argues that economic theory can give additional valuable input to green marketing. A first important step for marketing according to the economic approach is to identify how consumers perceive the products' cost and benefits. In this paper, cost and benefits are differentiated into different categories, which may help companies in finding their products' value added. In a second phase, companies can focus on stressing product advantages rather than on overcoming barriers. Using the example of green clothes this contribution shows what such a marketing strategy can look like. Copyright © 2001 John Wiley & Sons, Ltd and ERP Environment
Intimate partner stalking and femicide: urgent implications for women's safety
Authors: McFarlane Judith , Campbell Jacquelyn C. , Watson Kathy
Published: August 2002

This study describes the type and extent of intimate partner stalking and threatening behaviors that occurred within 12 months prior to a major assault or attempted or actual partner femicide and specifies which behaviors were associated with an increased risk of potential or actual lethality. The design was a ten‐city case–control study of 821 women: 384 abuse victims and 437 attempted or actual femicide informants. Data were derived using a 16‐item inventory. Logistic regressions, with adjustments for demographic variables, were used to identify the significant perpetrator behaviors associated with attempted/actual femicide. Women who reported the perpetrator followed or spied on them were more than twice as likely to become attempted/actual femicide victims. Threats by the perpetrator to harm the children if the woman left or did not return to the relationship place the woman at a ninefold increase in the risk of attempted/actual femicide. Conclusions are that certain stalking and threatening behaviors are strong risk factors for lethality, and women must be so advised. Copyright © 2002 John Wiley & Sons, Ltd.
Horizontal and vertical integration of academic disciplines in the medical school curriculum
Authors: Vidic Branislav , Weitlauf Harry M.
Published: August 2002

A rapid expansion of new scientific information and the introduction of new technology in operative and diagnostic medicine has marked the last several decades. Medical educators, because of and parallel to these developments, initiated a search for a more effective system of presenting core material to medical students. The new educational trends, although varying somewhat from one institution to another, concentrated on the following pedagogical shifts: 1) expansion of conceptual presentation of material at the expense of detail‐oriented education; 2) amplification of an integrated approach, as opposed to subject‐oriented instruction; 3) scheduling of elective courses to compliment required courses in the curriculum; and 4) institution of small group instruction (i.e., problem‐based learning) to actively involve students in the educational process and to develop deductive reasoning based on clinical cases. The future pedagogical system in medical schools will most likely be a combination of “classical” presentation of material combined with concept‐oriented, subject‐integrated and small group instruction based on either hypothetical or real clinical cases. It is imperative for the success of the new curriculum, however, that certain criteria are satisfied: 1) reorganize basic science departments to determine course ownership; 2) establish a reward system for teaching faculty; and 3) establish new course objectives. Clin. Anat. Month:233–235, 2002. © 2002 Wiley‐Liss, Inc.
Maintaining front‐line workers in child protection: a case for refocusing supervision
Authors: Gibbs Judy A.
Published: August 2002

This paper suggests that a refocusing of supervision be seen as one possible strategy for lowering high attrition rates among child protection workers. Research findings from a qualitative study undertaken in two rural regions in the State of Victoria, Australia illustrate that the current model of supervision gives insufficient attention to the emotional intrusiveness of the work, to building resilience in workers and the implications of adult learning theory. An argument is made for a refocusing of supervision which moves beyond task, encompassing much of what is already known from the supervision literature. The supervisor can be thought of as a messenger who must send and receive messages to and from workers. What message is sent and how supervisors respond to what they receive are critical if the objective is to retain front‐line staff. Critically, the supervisor must affirm both the merit and necessity of exploring the impact of feelings and thoughts on action and perception. A key message for supervisors to deliver is the value of individual workers to the organization, leading to increased self‐esteem and self‐efficacy. The paper also argues for adult learning via reflective supervision. This change to supervision involves a fundamental shift in organizational priorities and an acceptance throughout of the impact of anxiety on all those working in this field. Copyright © 2001 John Wiley & Sons, Ltd.
Life on the streets: the experiences of community researchers in a study of prostitution
Authors: Boynton Petra M.
Published: August 2002

Whilst community psychology has encouraged discussion of research processes and ways to improve studies, certain issues within community‐based research (particularly on sex topics) still require attention. This paper outlines a community study of women involved in street prostitution, and discusses issues arising from this research. By using extracts from the research report, along with researcher's diaries and taped conversations, the paper raises points for discussion concerning the process of sex‐research. For instance, training peer researchers, incorporating sex‐workers in research strategies, producing tangible results, and ethical issues. Strategies to improve research in general are then suggested. Copyright © 2002 John Wiley & Sons, Ltd.
Does contact with lesbians and gays lead to friendlier attitudes? a two year longitudinal study
Authors: Anderssen Norman
Published: August 2002

Attitudes toward and contact with lesbian women and gay men were assessed through questionnaires in a 2‐year follow‐up study in a cohort of 511 Norwegian participants, 19 years of age at first data collection. A substantial proportion reported negative attitudes at baseline (between 39 and 63%) and two years later (between 32 and 59%), females being more positive than males. A majority reported having no contact with lesbian women or gay men either at 19 years of age (66–79%) or 2 years later (51–62%). Longitudinally, the measures were moderately stable (Pearson's r 0.43–0.73), and contact change was positively related to attitude change. Also, attitude change positively predicted contact change. The findings imply that efforts to reduce stigma associated with homosexuality are still needed among young people in Norway. Copyright © 2002 John Wiley & Sons, Ltd.
Effect of vitamin C and zinc on osmotic fragility and lipid peroxidation in zinc‐deficient haemodialysis patients
Authors: Candan Ferda , Gültekin Füsun , Candan Ferhan
Published: August 2002

Peroxidation of the membrane lipid structure of red blood cell leads to haemolysis and anaemia in haemodialysis patients. Dietary constituents of antioxidant vitamins and trace elements may play an important role in protecting against oxidant damage. In this study, the effects of supplementation of vitamin C and zinc on osmotic fragility and lipid peroxidation of erythrocytes were investigated in 34 zinc‐deficient haemodialysis patients. Sixteen sex‐ and age‐matched normal volunteers acted as controls. Patients were randomized to receive vitamin C (250 mg day−1), zinc (20 mg day−1) or a placebo treatment for 3 months. The levels of vitamin C, zinc, malondialdehyde (MDA) and osmotic fragility were measured initially and 3 months after supplementation. Mean serum concentration of vitamin C and zinc increased significantly in the groups at the end of the respective study periods. Supplementation with vitamin C and zinc improved osmotic fragility, and decreased the level of MDA in the groups, but some side‐effects (i.e. nausea, vomiting, fever, muscle pain, weakness) were observed during the zinc treatment. The results showed that the supplementation of both treatments decreased osmotic fragilty and MDA in zinc‐deficient haemodialysis patients. However, vitamin C treatment was found to be safer than zinc supplementation. Copyright © 2001 John Wiley & Sons, Ltd.
State Policy and Postsecondary Enrollment Options: Creating Seamless Systems
Authors: Boswell Katherine
Published: August 2002

Hundreds of thousands of secondary students across the nation—as many as 10 percent of high school students in some states—are participating in various forms of postsecondary enrollment options. State policymakers are considering a range of new policy options to accelerate student educational progress in light of projected demands for access to public colleges and universities by the "baby boom echo."
Directional atherectomy prior to stenting in bifurcation lesions: A matched comparison study with stenting alone
Authors: Karvouni Evangelia , Di Mario Carlo , Nishida Takahiro , Tzifos Vaios , Reimers Bernhard , Albiero Remo , Corvaja Nicola , Colombo Antonio
Published: August 2002

The ideal catheter‐based intervention for treatment of coronary lesions at bifurcation site still has to be defined. The aim of the study was to assess the acute and long‐term outcome after treatment of bifurcation lesions with directional atherectomy (DCA) and stenting in comparison with stenting alone. Thirty‐one consecutive patients treated for bifurcation coronary lesions (62 lesions) with DCA and stenting in at least one branch (DCA group) were compared with a matched group of 31 patients with bifurcation coronary lesions (62 lesions) treated with stenting alone in at least one branch (non‐DCA group). Procedural success was 87.1% in the DCA group compared with 100% in the non‐DCA group (P = 0.03). In‐hospital major adverse cardiac events (MACE) occurred only in the DCA group (12.9% vs. 0%, P = 0.03), mainly non–Q‐wave myocardial infarction. After the procedure, minimum lumen diameter (MLD) and acute gain were significantly greater (P = 0.004 and P = 0.05, respectively) and % diameter stenosis was significantly lower (P = 0.05) in the main branch in the DCA group. At follow‐up angiogram, MLD in the main branch was still significantly greater in the DCA group compared to the non‐DCA group (2.31 vs. 1.65, respectively, P = 0.04), with no significant difference in late loss and loss index between the two groups. Restenosis rate was 28.8% in the DCA group vs. 43.5% in the non‐DCA group (P = 0.13). The incidence of follow‐up MACE was 29% in the DCA group compared with 48.4% in the non‐DCA group, mainly due to target lesion revascularization. In conclusion, treatment of bifurcation coronary lesions with DCA and stenting was associated with greater acute gain after the procedure and greater MLD at follow‐up in the main branch compared with stenting alone. Procedural myocardial infarction was more frequent in the DCA group. Restenosis rates and follow‐up MACE were lower following DCA and stenting, without reaching any statistical significance. Cathet Cardiovasc Intervent 2001;53:12–20. © 2001 Wiley‐Liss, Inc.
Evolution in the practice of primary angioplasty: Effect of adjunctive coronary stenting and glycoprotein IIb/IIIa inhibitors on long‐term outcomes
Authors: Kalaria Vijay G. , Chaudhary Imran , Jacobson Seth , Pomerantz Richard M. , Cove Christopher J. , Ling Frederick S. , Cutlip Donald E.
Published: August 2002

Primary angioplasty strategies have evolved dramatically, including increasing adjunctive use of stents and glycoprotein (GP) IIb/IIIa inhibitors. The purpose of this study was to examine the specific effects of these adjunctive therapies on long‐term outcomes after primary angioplasty. From 1996 to 1998, 257 unselected, consecutive patients underwent primary PTCI at our institution. In‐hospital mortality was 5.4% (2.9% for patients without cardiogenic shock). The remaining 243 patients were followed for 2.0 ± 0.7 years. Adjunctive stenting was associated with increased late mortality (8.7% vs. 2.3%, P = 0.02). GP IIb/IIIa inhibitors were associated with reduced late mortality among patients receiving stents (6.9% vs. 21.4%, P = 0.07), but not in those patients treated with balloon angioplasty alone (2.9% vs. 0%, P > 0.20). Coronary stenting remained a significant predictor of late mortality (hazard ratio 5.6, 95% CI 1.5–21.2) after adjustment for other established risk factors. In this unselected series, adjunctive coronary stenting was associated with higher late mortality among hospital discharge survivors. Concomitant GP IIb/IIIa inhibitors partially corrected for this increase. These results are limited by the small sample size and retrospective design of this study. Additional long‐term studies are required to test these findings and evaluate for possible mechanisms. Cathet Cardiovasc Intervent 2001;54:327–332. © 2001 Wiley‐Liss, Inc.
Stereospecificity and stereoselectivity of flobufen metabolic profile in male rats in vitro and in vivo: Phase I of biotransformation
Authors: Wsól Vladimír , Král Radim , Skálová Lenka , Szotáková Barbora , Trejtnar František , Flieger Miroslav
Published: August 2002

Flobufen (F) is the original nonsteroidal antiinflammatory drug (NSAID) containing two enantiomers. The aim of this investigation was to elucidate the biotransformation pathway of F at chiral level in phase I of biotransformation. Stereoselectivity and stereospecificity of the respective enzymes were studied in male rats in vitro (microsomal and cytosolic fractions, hepatocytes suspension) and in vivo. The rac‐F, (+)‐R‐F and (−)‐S‐F were used as substrates. Amounts of F enantiomers, 4‐dihydroflobufen diastereoisomers (DHF) and other metabolites (M‐17203, UM) were determined with a chiral HPLC method in two chromatographic runs on R,R‐ULMO and allyl‐terguride bonded columns. Stereoselective biotransformation of the two enantiomers of F was observed at all tested levels and significant bidirectional chiral inversion of enantiomers of F was observed in hepatocytes. Mean enantiomeric ratios of F concentrations (S‐/R‐), after rac‐F incubations, ranging from 1.09 in cytosolic fraction to 18.23 in hepatocytes. Stereospecificity of the respective F reductases was also observed. (2R;4S)‐DHF and (2S;4S)‐DHF are the principal metabolites of F in microsomes and hepatocytes. Neither DHF diastereoisomers nor M‐17203 were found in cytosolic fraction. Only the nonchiral metabolite, M‐17203, was found in all urine and feces samples after oral administration of F. Chirality 13:754–759, 2001. © 2001 Wiley‐Liss, Inc.
Application of L‐threonine aldolase‐catalyzed reaction to the preparation of protected 3R,5R‐dihydroxy‐L‐homoproline as a mimetic of idulonic acid
Authors: Miura Tsuyoshi , Kajimoto Tetsuya
Published: August 2002

A facile synthesis of 3R,5R‐dihydroxy‐L‐homoproline as idulonic acid mimic, of which the carboxyl and 3‐hydroxyl groups were protected, was attained using L‐threonine aldolase‐catalyzed reaction. Idulonic acid is a key acidic sugar of the b‐FGF binding domain in heparin and heparan sulfate. Moreover, the synthetic precursor of N‐acetyl‐4‐deoxy‐D‐mannosamine, which is a potent inhibitor of NeuAc synthase, was prepared from the side product of the enzymatic aldol condensation. Chirality 13:577–580, 2001. © 2001 Wiley‐Liss, Inc.
Regulation of flagellar dynein by the axonemal central apparatus
Authors: Smith Elizabeth F.
Published: August 2002

Numerous studies indicate that the central apparatus, radial spokes, and dynein regulatory complex form a signaling pathway that regulates dynein activity in eukaryotic flagella. This regulation involves the action of several kinases and phosphatases anchored to the axoneme. To further investigate the role of the central apparatus in this signaling pathway, we have taken advantage of a microtubule‐sliding assay to assess dynein activity in central apparatus defective mutants of Chlamydomonas. Axonemes isolated from both pf18 and pf15 (lacking the entire central apparatus) and from pf16 (lacking the C1 central microtubule) have reduced microtubule‐sliding velocity compared with wild‐type axonemes. Based on functional analyses of axonemes isolated from radial spokeless mutants, we hypothesized that inhibitors of casein kinase 1 (CK1) and cAMP dependent protein kinase (PKA) would rescue dynein activity and increase microtubule‐sliding velocity in central pairless mutants. Treatment of axonemes isolated from both pf18 and pf16 with DRB, a CK1 inhibitor, but not with PKI, a PKA inhibitor, restored dynein activity to wild‐type levels. The DRB‐induced increase in dynein‐driven microtubule sliding was inhibited if axonemes were first incubated with the phosphatase inhibitor, microcystin. Inhibiting CK1 in pf15 axonemes, which lack the central pair as well as PP2A [Yang et al., 2000: J. Cell Sci. 113:91–102], did not increase microtubule‐sliding velocity. These data are consistent with a model in which the central apparatus, and specifically the C1 microtubule, regulate dynein through interactions with the radial spokes that ultimately alter the activity of CK1 and PP2A. These data are also consistent with localization of axonemal CK1 and PP2A near the dynein arms. Cell Motil. Cytoskeleton 52:33–42, 2002. © 2002 Wiley‐Liss, Inc.
Inducing precocious anaphase in cultured mammalian cells
Authors: Canman Julie C. , Salmon E.D. , Fang Guowei
Published: August 2002

The spindle checkpoint, which prevents anaphase onset upon spindle damage or incorrect chromosome alignment, presents a problem for experimental analysis of protein function in anaphase and cytokinesis. This is because the functional disruption of many proteins before anaphase onset can activate this checkpoint, preventing anaphase and subsequent cell cycle events. This paper compares new and old methods of overriding the spindle checkpoint in prometaphase mammalian tissue culture cells. Cell Motil. Cytoskeleton 52:61–65, 2002. © 2002 Wiley‐Liss, Inc.
A healthy understanding of intraflagellar transport
Authors: Sloboda Roger D.
Published: August 2002

A microtubule‐dependent motility process called intraflagellar transport (IFT) occurs beneath the plasma membrane of cilia and flagella. IFT was first observed in Chlamydomonas, and orthologs of some of the polypeptides involved in IFT have recently been identified in other organisms, including C. elegans and the mouse. In addition to a role in the assembly and maintenance of cilia and flagella, evidence is reviewed here that indicates defects in the process of IFT may be related to problems with human health. Moreover, recent data suggest the possibility of two new roles for IFT in cell function. The first is in transcriptional control of the genes encoding ciliary and flagellar proteins. IFT could provide a mechanism whereby the cell senses the presence or absence of its cilia or flagella and responds by turning on gene transcription resulting in replacement of the missing organelle. The second role is in signal transduction, whereby cilia act as sensors of the external cellular environment and transduce information about the surroundings into intracellular signals that are sent via IFT to the cell body, thus inducing an appropriate cellular response to the environment. Cell Motil. Cytoskeleton 52:1–8, 2002. © 2002 Wiley‐Liss, Inc.
Coexpression of ephrin‐Bs and their receptors in colon carcinoma
Authors: Liu Wenbiao , Ahmad Syed A. , Jung Young D. , Reinmuth Niels , Fan B.S. Fan , Bucana Corazon D. , Ellis Lee M.
Published: August 2002

BACKGROUND

The erythropoietin‐producing hepatoma amplified sequence (Eph) family is the largest subfamily of receptor tyrosine kinases (RTKs). The Ephs (receptors) bind to specific cell‐bound ligands, called ephrins. The binding of this ligand‐receptor system is dependent on cell‐cell interactions. The ephrin‐Eph system is important in embryologic development and differentiation of the nervous and vascular systems. In the current study, the authors hypothesized that ephrins may play a role in the growth and development of colon carcinoma and may be expressed differentially in normal and malignant colonic tissues.

METHODS

Reverse transcriptase‐polymerase chain reaction (RT‐PCR), Northern Blot analyses, and immunohistochemistry were used to examine 11 colon carcinoma cell lines and 20 human colon carcinoma specimens with adjacent uninvolved mucosa for the expression of EphB and ephrin‐B family members.

RESULTS

EphB2, EphB3, and EphB4 mRNA expression and ephrin‐B2 mRNA expression was detected in all the cell lines and colon carcinoma specimens examined. Immunohistochemical analysis showed that ephrin‐B2 had higher expression in the colon carcinoma specimens studied than in adjacent normal mucosa. Ephrin‐B2 and EphB4 most frequently were expressed on the luminal surface of colon carcinoma epithelium.

CONCLUSIONS

The results of the current suggest that the ephrin‐Bs are expressed differentially in colon carcinoma and normal mucosa specimens and thus may play a role in the progression of colon carcinoma. Further studies are necessary to determine the functional role of ephrin‐Bs in colon carcinoma angiogenesis and growth. Cancer 2002;94:934–9. © 2002 American Cancer Society.

DOI 10.1002/cncr.10122
Lack of significant differences in the corrected activity of lysophospholipase D, producer of phospholipid mediator lysophosphatidic acid, in incubated serum from women with and without ovarian tumors
Authors: Tokumura Akira , Tominaga Kyoko , Yasuda Katsuhiko , Kanzaki Hideharu , Kogure Kentaro , Fukuzawa Kenji
Published: August 2002

BACKGROUND

Several studies have shown that lysophosphatidic acid (LPA), a phospholipidic chemical mediator, is relevant to the pathogenesis of ovarian carcinoma. Higher plasma levels of LPA have been reported in patients with ovarian carcinoma than in healthy patients, and LPA is known to activate ovarian carcinoma cells. To determine the reason for the increased plasma LPA levels in ovarian carcinoma patients, we compared the activities of serum lysophospholipase D, a novel LPA‐producing metallo‐enzyme, in healthy volunteers, patients with benign ovarian tumor, and patients with ovarian carcinoma.

METHODS

Lysophospholipase D activity was assessed by measuring the percentage conversion of [14C]palmitoyl‐lysophosphatidylcholine (LPC) added to human serum. The apparent enzyme activities were corrected based on the serum levels of palmitoyl‐LPC determined by gas‐liquid chromatography after its purification and conversion to fatty acid methyl esters.

RESULTS

The apparent activity of lysophospholipase D in serum preparations from four patients with ovarian carcinoma at Stage IV was significantly higher than those from five healthy subjects, five patients with benign ovarian tumors, and fourteen patients with ovarian carcinoma at Stages I (n = 5), II (n = 4), and III (n = 5). The serum levels of LPC, an endogenous substrate of lysophospholipase D, in ovarian carcinoma patients were less than those in patients with benign ovarian tumors. There were no significant differences in the corrected lysophospholipase D activity for the LPC levels in healthy women, patients with benign ovarian tumors, and patients with ovarian carcinoma at various stages.

CONCLUSIONS

The current results suggest that lysophospholipase D is not associated with the elevated plasma levels of LPA in ovarian carcinoma patients previously reported, although only a limited number of patients were analyzed. Cancer 2002;94:141–51. © 2002 American Cancer Society.
Influences of the lysosomal associated membrane proteins (Lamp‐1, Lamp‐2) and Mac‐2 binding protein (Mac‐2‐BP) on the prognosis of pancreatic carcinoma
Authors: Künzli Beat M. , Berberat Pascal O. , Zhu Zhaowhen W. , Martignoni Marcus , Kleeff Jörg , Tempia‐Caliera Adrien A. , Fukuda Minoru , Zimmermann Arthur , Friess Helmut , Büchler Markus W.
Published: August 2002

BACKGROUND

Lamps and Mac‐2‐BP are ligands of galectin‐3, and they were suggested to influence tumor proliferation and metastasis formation. The authors studied the expression of Lamp‐1, Lamp‐2, and Mac‐2‐BP in pancreatic carcinoma and evaluated their influence on patient prognosis.

METHODS

Northern blot analysis, in situ hybridization, and immunohistochemistry were performed in 12 normal and 28 pancreatic carcinoma tissue samples and in pancreatic carcinoma cell lines. The molecular findings in the tumor samples were correlated with the prognosis and histopathologic tumor characteristics. In addition, in Lamp‐1 transfected CAPAN‐1 pancreatic carcinoma cells, cell proliferation was analyzed.

RESULTS

Lamp‐1, Lamp‐2, and Mac‐2‐BP were overexpressed in 61% (1.6‐fold increase, not significant), 71% (3.0‐fold increase, P < 0.01), and 93% (5.6‐fold increase, P < 0.01) of the pancreatic carcinoma samples. Lamp‐1 and Lamp‐2 immunoreactivity was present at the luminal side of the ductal carcinoma cells whereas Mac‐2‐BP immunoreactivity was diffusely spread over the whole cytoplasm and the nucleolus of ductal carcinoma cells. Correlation of the molecular data with clinical patient parameters revealed that patients whose tumors exhibited high Lamp‐1 mRNA expression lived significantly longer (median, 17 months) after tumor resection than patients whose tumors exhibited low to moderate Lamp‐1 mRNA levels (median, 8 months; P < 0.02). No relation between Lamp‐2 and Mac‐2‐BP mRNA expression and any of the histopathologic parameters was found. Lamp‐1 transfected CAPAN‐1 cells showed decreased cell growth compared with the nontransfected cells.

CONCLUSIONS

Lamp‐1 might influence local tumor progression rather than the formation of tumor metastasis in pancreatic carcinoma, whereas Mac‐2‐BP and Lamp‐2 seem to have little influence on these parameters in pancreatic carcinoma. Cancer 2002;94:228–39. © 2002 American Cancer Society.
Very early detection of RET proto‐oncogene mutation is crucial for preventive thyroidectomy in multiple endocrine neoplasia type 2 children
Authors: Sanso Gabriela E. , Domene Horacio M. , Garcia Rudaz Maria C. , Pusiol Eduardo , de Mondino Ana K. , Roque Maria , Ring Alejandro , Perinetti Hector , Elsner Boris , Iorcansky Sonia , Barontini Marta
Published: August 2002

BACKGROUND

Multiple endocrine neoplasia type 2 (MEN 2) is an inherited disease caused by germline mutations in the RET proto‐oncogene, and is responsible for the development of endocrine neoplasia. Its prognosis is dependent on the appearance and spread of medullary thyroid carcinoma (MTC). Relatives at risk can be identified before clinical or biochemical signs of the disease become evident.

METHODS

Twenty‐one families with MEN 2 (16 families with MEN 2A and 5 families with MEN 2B) were studied. Peripheral blood DNA was amplified by polymerase chain reaction. DNA sequence or restriction enzyme analysis was performed to detect mutations of RET proto‐oncogene exons 10, 11, and 16. Molecular analysis was carried out in all index patients as well as in 98 relatives of MEN 2A patients (60 juveniles, ages 6 months to 21 years, and 38 adults, ages 22 to 81 years) and in 13 relatives (6 juveniles ages 10 to 21 years, and 7 adults ages 41 to 66 years) from MEN 2B families.

RESULTS

Molecular studies showed a mutation at codon 634, exon 11 in all MEN 2A patients. All MEN 2B patients showed an ATG to ACG (Met918Thr) mutation. In MEN 2A families, 42 out of 98 relatives were affected. Total thyroidectomy was performed in 18 juvenile carriers ages 17 months to 21 years. Histopathologic studies of the glands revealed parafollicular cell (C‐cell) hyperplasia in all of these carriers, medullary thyroid carcinoma in 15 carriers, and only one carrier with lymph node metastases.

CONCLUSIONS

The consistent finding of C‐cell disease in all the juvenile carriers who underwent preventive thyroidectomy emphasizes the relevance of early screening in children at risk of developing MTC. The presence of MTC in the specimen of prophylactic thyroidectomy from a 17 month old girl highlights the importance of thyroidectomy as soon as the molecular diagnosis is confirmed. Cancer 2002;94:323–30. © 2002 American Cancer Society.
Central nervous system relapse in adults with acute lymphoblastic leukemia
Authors: Surapaneni Usha R. , Cortes Jorge E. , Thomas Deborah , O'Brien Susan , Giles Francis J. , Koller Charles , Faderl Stefan , Kantarjian Hagop
Published: August 2002

BACKGROUND

Recurrence of acute lymphoblastic leukemia (ALL) in the central nervous system (CNS) in adults is considered a poor prognostic feature. Few reports have analyzed this issue. The objective of this study was to analyze the experience with CNS recurrence in adult patients with ALL.

METHODS

Between December 1982 and April 2000, the records of 527 consecutive, newly diagnosed patients with ALL who were treated at the M. D. Anderson Cancer Center were reviewed. Patients were treated with one of two published regimens: vincristine, doxorubicin, and dexamethasone (VAD; n = 237 patients) or hyperfractionated cyclophosphamide plus VAD (hyper‐CVAD) (n = 290 patients). Among 439 patients (83%) who achieved complete remission (CR), 32 patients (7%) had a CNS recurrence. All patients had received CNS prophylaxis tailored to the risk of CNS recurrence with systemic chemotherapy with or without intrathecal (IT) chemotherapy; none received cranial radiotherapy for prophylaxis. The records of these patients were reviewed.

RESULTS

The median age at the time of diagnosis was 33 years. The median CR duration prior to CNS recurrence was 36 weeks (range, 2–185 weeks). Three groups were identified: 1) patients with an isolated CNS recurrence (n = 17 patients), 2) patients with CNS recurrences after bone marrow (BM) recurrence (n = 8 patients), and 3) patients with simultaneous CNS and BM recurrences (n = 7 patients). IT chemotherapy was effective in achieving a CNS CR in 30 patients (94%), but 10 patients (31%) had a second CNS recurrence. The median survival from CNS recurrence was 6 months; 28% of patients were alive at 1 year, and 6% of patients were alive at 4 years.

CONCLUSIONS

Adults with ALL and CNS recurrences have a poor prognosis despite effective IT chemotherapy. Future studies should investigate better approaches in the treatment of these patients to improve their long‐term survival. Effective CNS prophylaxis remains the single best approach for treating patients with CNS leukemia. Cancer 2002;94:773–9. © 2002 American Cancer Society.

DOI 10.1002/cncr.10265
Presence of active gelatinases in endometrial carcinoma and correlation of matrix metalloproteinase expression with increasing tumor grade and invasion
Authors: Di Nezza Lisa A. , Misajon Aileen , Zhang Jin , Jobling Tom , Quinn Michael A. , Östör Andrew G. , Nie Guiying , Lopata Alex , Salamonsen Lois A.
Published: August 2002

BACKGROUND

The actions of the extracellular‐matrix degrading enzymes, matrix metalloproteinases (MMPs), are implicated in tumorigenesis. The cellular localization of MMP‐2, MMP‐9, membrane type 1 (MT1)‐MMP, tissue inhibitors of metalloproteinases (TIMPs) 1‐3, and the presence of active gelatinases were investigated in endometrial carcinoma.

METHODS

Endometrial carcinomas were grouped according to histologic grade (Grades 1‐3), depth of myometrial invasion (0, < 50%, > 50%) and the presence of vascular/lymphatic invasion. Twenty‐nine endometrial carcinoma biopsies were investigated immunohistochemically to determine the tissue localization of MMP‐2 (gelatinase A), MMP‐9 (gelatinase B), MT1‐MMP, and TIMPs 1‐3. In situ hybridization was performed to localize MMP‐2 and MMP‐9 mRNA. The presence of active gelatinases was assessed using in situ zymography.

RESULTS

Epithelial tumor cells were the main site of MMP‐2, MMP‐9, and MT1‐MMP protein. Variable stromal cell localization was also observed, particularly in areas adjacent to tumor nests. Semiquantitative analysis revealed increases in MMP‐9 and MMP‐2 but not MT1‐MMP staining scores in tumor epithelial cells in the transition from histologic Grade 1 to Grades 2 and 3. Matrix metalloproteinase‐9 and MT1‐MMP staining scores in tumor cells were significantly associated with the presence of myometrial invasion and vascular/lymphatic invasion, while MMP‐2 did not correlate with these factors. In addition, MT1‐MMP was co‐localized with MMP‐2, supporting its role in the activation of proMMP‐2. Tumor cells from all histologic grades stained intensely for TIMP‐2 and TIMP‐3 proteins, while variable stromal staining was observed. In Grade 1 carcinomas TIMP‐1 was predominantly immunolocalized to the stromal compartment with variable tumor cell localization being observed in Grades 2 and 3 carcinomas. Matrix metalloproteinase‐9 and MMP‐2 mRNAs were predominantly observed in tumor epithelial cells as well as in the stroma to varying degrees. In situ zymography revealed active forms of gelatinases at the cellular surface and in association with tumor epithelial cells within endometrial carcinoma tissues.

CONCLUSIONS

These data suggest that increasing expression of MMPs and endometrial carcinoma progression are closely related. Active gelatinases are present in endometrial carcinoma, resulting in alterations to the microenvironment that promote tumor invasion and metastasis. Cancer 2002;94:1466–75. © 2002 American Cancer Society.

DOI 10.1002/cncr.10355
Trimodality treatment in Stage III nonsmall cell lung carcinoma
Authors: Broermann Petra , Junker Klaus , Brandt Burkhard H. , Heinecke Achim , Freitag Lutz , Klinke Folker , Berdel Wolfgang E. , Thomas Michael
Published: August 2002

BACKGROUND

In a trimodality treatment approach for Stage III nonsmall cell lung carcinoma (NSCLC), the prognostic impact of the ras mutation status in resection specimens was evaluated.

METHODS

Forty patients with Stage III NSCLC underwent tumor resection after neoadjuvant treatment with two cycles of chemotherapy (ifosfamide, carboplatin, and etoposide) and subsequent twice‐daily radiotherapy (45 grays [Gy]; 2 × 1.5 Gy/day) with concurrent carboplatin and vindesine. Assessment of K‐ras codon 12 mutation status was performed in the paraffin embedded resection specimens by a two‐step polymerase chain reaction followed by restriction fragment length polymorphism analysis.

RESULTS

K‐ras mutation status could be assessed in 28 cases. A K‐ras codon 12 point mutation was found in 13 of 28 resection specimens (46%). The mutation was found independently of gender, age, tumor stage, and clinical response status and occurred more frequently in adenocarcinomas. Even after complete resection, the presence of a K‐ras mutation was a significant predictor for a poor progression free survival (P = 0.005).

CONCLUSIONS

These data suggest that further evaluation of the K‐ras codon 12 mutation status in trials on neoadjuvant and adjuvant therapy is warranted. This may contribute to the identification of stratification variables for future treatment approaches. Cancer 2002;94:2055–62. © 2002 American Cancer Society.

DOI 10.1002/cncr.10387
Cell types and response properties of neurons in the ventral division of the medial geniculate body of the rabbit
Authors: Cetas Justin S. , Price Robin O. , Velenovsky David S. , Crowe Jennifer J. , Sinex Donal G. , McMullen Nathaniel T.
Published: August 2002

Although there is evidence for multiple classes of thalamic relay neurons in the auditory thalamus, correlative anatomical and physiological studies are lacking. We have used the juxtacellular labeling technique, in conjunction with Nissl, Golgi, and immunocytochemical methods, to study the morphology and response properties of cells in the ventral division of the medial geniculate body of the rabbit. Single units in the ventral division of the medial geniculate body (MGV) were characterized extracellularly with monaural and binaural tone and noise bursts (100‐ to 250‐msec duration). Characterized units were filled with biocytin and visualized with an antibody enhanced diaminobenzidine reaction. A total of 31 neurons were physiologically characterized and labeled with the juxtacellular technique. Labeled neurons were fully reconstructed from serial sections by using a computer microscope system. Three subregions of the rabbit MGV were identified, each characterized by differences in Nissl architecture, calcium‐binding protein expression, and by the dendritic orientation of tufted relay neurons. In general, the dendritic fields of relay neurons were closely aligned with the cellular laminae. Qualitative and quantitative analyses revealed two types of presumptive relay neurons within the MGV. Type I cells had thick dendrites with a greater total volume and morphologically diverse appendages compared with the Type II cells whose dendrites were thin with a moderate number of small spines. Both classes were acoustically responsive and exhibited a variety of response patterns, including onset, offset, and sustained responses. In terms of binaural characteristics, most (ca. 53%) labeled neurons were of the EE type, with the remaining cells classified as EO (27%) or EI (20%) response types. Two types of presumptive interneurons were also seen: bipolar neurons with large dendritic fields and a small neurogliaform variety. Cell types and dendritic orientation within the MGV are discussed in terms of the physiological organization of the rabbit auditory thalamus. J. Comp. Neurol. 445:78–96, 2002. © 2002 Wiley‐Liss, Inc.