bluuebunny/crossref_metadata_2025 huggingface dataset:
Extending studies of collaborative cognition by way of care giving situations
Authors: Goodnow Jacqueline J. , Lawrence Jeanette A. , Ryan Jacinta , Karantzas Gery , King Kylie
Published: July 2002

Care giving situations contain several features that offer opportunities for expanding the way that collaborative cognition is conceptualised and explored. These features are the presence of several possible contributors, more than one kind of change in participation, distinctions drawn among parts of a task, and differences in understanding based on interests. All represent departures from the traditional focus on dyads, tasks that emphasise one kind of change only, single problems, and differences in competence or expertise. All are also features likely to be found in everyday problem solving. Study 1 focuses on family contributions, based on reports by care givers about their current situation and their preferences for the involvement of other family members. Study 2 presents a standard family scenario and focuses on the views held by care givers, older adults, and community nurses about the reasonableness of various changes in participation. Results are discussed in terms of the ways situations such as care giving can help build a richer picture of collaborative cognition, one that is applicable to a variety of tasks and to all parts of the life span.
Personality Development between 1 and 8 Years of Age in Swedish Children with Varying Child Care Experiences
Authors: Wessels Holger , Lamb Michael E. , Hwang C. Philip , Broberg Anders G.
Published: July 2002

In this longitudinal study, 122 Swedish children were followed from an initial assessment at 16 months until they were 8.5 years of age. Parental ratings of field independence, ego-resilience, and ego-control were obtained using the California Child Q-set when the children averaged 28, 40, 80, and 101 months of age. These assessments of personality were then examined in the context of contrasting child care experiences. There was virtually no difference between the developmental trajectories of the children in home care and in centre-based day care. In contrast, ego-undercontrol decreased less, whereas ego-resilience and field independence increased less, in children who attended family day care than in the children in either centre care or exclusive parental care. The quality of home and out-of-home care, as well as socioeconomic status and family background, did not moderate or qualify these effects of the type of care experienced. The results are discussed in relation to previous work on the effects of early out-of-home care on child development.
Similarities and Differences in Mothers’ and Observers’ Descriptions of Attachment Behaviours
Authors: Tarabulsy George M. , Avgoustis Effie , Phillips Jennifer , Pederson David R. , Moran Greg
Published: July 2002

This report concerns the similarities and differences between maternal and observer Attachment Q-Set (AQS) security scores for 41 preterm and 38 fullterm infants. The following variables were assessed: maternal sensitivity (8 months); parental stress (8 months); mother- and observer-derived AQS measures of attachment security and dependency (12 months); and infant temperament (18 months). “Strange situations” were recorded at 18 months. Previous analyses of these data had shown that although mother and observer AQS security scores were correlated ( r = .55), only observer scores converged with the strange situation. Two additional series of analyses were carried out. The first revealed that AQS security scores of sensitive mothers, but not those of less sensitive mothers, are correlated with those of observers. Maternal sensitivity was unrelated to the lack of correspondence between mothers’ AQS scores and strange situation classification. Second, a factor analysis of the above variables revealed that while observers’ sensitivity and attachment security scores and mothers’ security scores loaded on to a Relationship Security factor, mothers’ security ratings also loaded on to an Infant Fussiness factor. This finding suggests that although both mothers and observers focus on items indicative of security when completing the AQS, mothers may place an additional emphasis on fussiness-related items. Further support for this suggestion was found in an analysis of covariance. When variance attributed to fussiness-related items of the AQS was partialed out of the mother-derived security score, the residual mapped on to the secure/nonsecure distinction in the strange situation. However, fussiness was unrelated to the mother-observer AQS correlation, indicating that different sources of variance may be involved in the mother-observer AQS correlation and in the correspondence between mothers’ AQS scores and the strange situation.
Control Judgements across the Lifespan
Authors: Grob Alexander , Little Todd D. , Wanner Brigitte
Published: July 2002

Given the inconsistent findings in the literature, we examined age-cohort differences in various personal control judgements over the lifespan ( N = 1623; ages 14-85). For three distinct life domains (personal, social, and societal), participants rated their amount of personal control, the goal importance, the degree of control striving, and, in comparison with same-aged peers, their relative control in each domain. Within this broad lifespan range, the developmental trends showed, as expected, different trajectories depending upon the type of control dimension and the nature of the life domain. The differential nature of these age-cohort trends have important implications for understanding the inconsistent findings reported in the literature, namely, that depending on the nature of the life domain, the type of control dimension, and the age range tested, the trajectories can either increase, decrease, or remain stable. These trends are discussed with reference to various metatheoretical perspectives on lifespan development and control-related judgements.
Changes in primary boys' and girls' attitudes to school and science during a two‐year science in‐service programme
Authors: Jarvis Tina , Pell Anthony
Published: July 2002

A pupils' attitude test was developed to monitor change over the two years of a primary science in‐service programme. It explores pupils' views about school in general so that changes in science attitudes can be contextualized within the whole school experience; feelings towards science experiments; and perceptions of real‐world science. The test was piloted with schools outside the in‐service programme. Approximately 2000 pupils were tested in January 1999, September 1999 or January 2000. A random sample of children were retested in June 1999 and compared with a ‘control’ school. All the children in the project were tested again in July 2000. The results indicate a generally positive attitude towards coming to school, with particular preferences for work with computers and working together. As the pupils get older, their enthusiasm for science falls as they find it less difficult or demanding. This relationship appears to be broken by teacher in‐service to some extent. Girls in particular also showed a greater enthusiasm for independent investigative science after the programme.
The prevalence and characteristics of injecting drug users in Australia*
Authors: MCALLISTER IAN , MAKKAI TONI
Published: July 2002

The prevalence of injecting drug use has increased significantly in Australia in recent years. Using the nationally representative 1998 National Drug Strategy Household Survey we estimate the size of the lifetime prevalence injecting drug use population at 302 710 people, compared to 67 474 in 1988. Those who have injected drugs in the previous year are estimated at 108 750 people in 1998. Amphetamines have displaced heroin as the most popular drug to inject among the general population. Those who have ever injected are more likely to be male, young, unemployed and to possess fewer educational qualifications. A variety of factors are discussed which may account for increases in injecting drug use.
Th1 and Th2 cytokine profile in patients with early onset periodontitis and their healthy siblings
Authors: Bártová Jiřina , Krátká-Opatrná Zuzana , Procházková Jarmila , Krejsa Otakar , Dušková Jana , Mrklas Lubor , Tlaskalová Helena , Cukrowská Božena
Published: July 2002

Early onset periodontitis (EOP) is a chronic inflammatory periodontal disease with a strong genetic link affecting individuals aged 17 to 25. In the familial studies we tested the hypothesis about the role of Th1 and Th2 cytokines in the pathogenesis of EOP disease. The study involved 6 individuals with EOP disease and their 6 siblings with healthy periodontium. Actinobacillus actinomycetemcomitans (A. a.), a bacterium typical for EOP, was detected in all people studied. Th1 and Th2 cytokine production was measured after in vitro stimulation. Peripheral blood mononuclear cells (PBMC) were isolated and cultivated for 24 h and 7 days with PWM, A. a. or Escherichia coli. The levels of IL–4, IFN‐gamma, IgA, IgG and IgM were measured by ELISA methods. After in vitro stimulation of PBMC, a significantly higher production of IL–4 and significantly lower production of IFN‐gamma were found in the group of patients compared with their healthy siblings. The increased level of IL–4 in patients was in good agreement with an increased level of IgM after stimulation of lymphocytes with E. coli. These results support Seymour’s hypothesis according to which patients with progressive disease primarily activate Th2 lymphocytes while non‐susceptible individuals activate Th1 lymphocytes.
Nicholson Versus Ingram on the History of Political Economy and a Charge of Plagiarism
Authors: Moore Gregory
Published: July 2002

In 1885 John Kells Ingram published a lengthy article on the history of political economy in the Encyclopaedia Britannica, and in 1888 he republished this same article, with only minor changes, as a book entitled The History of Political Economy. Ingram unashamedly interpreted the historical development of political economy from a Comtean variant of the historicist perspective, and, for this reason, these publications became extremely important for the methodological debate, known as the English Methodenstreit, then raging between the orthodox and historical economists. Although the historicist message dovetailed into Ingram's historical narrative was clearly contentious and polemical, the reviews of both versions of this history from either side of the conceptual divide were overwhelmingly positive. Two exceptions were damning anonymous newspaper reviews in The Scotsman: one in 1885 in response to the Encyclopaedia Britannica article, and another in 1888 in response to the book. It is apparent from an entry made in the diary of John Neville Keynes that the first of these reviews was written by Joseph Shield Nicholson (JNK, July 28, 1885, Add 7834), and since the articles are strikingly similar in tone, substance, and style, and since Nicholson wrote other reviews for The Scotsman, it is safe to assume that the second article was also written by Nicholson. The substance of Nicholson's critique can be reduced to two main accusations: first, that Ingram was neither qualified to write a history of political economy nor competent to comment on the methodological issues then under scrutiny, and second, that Ingram had brazenly plagiarized passages drawn from various German histories of political economy.
Inpatient Alcohol Treatment in a Private Healthcare Setting: Which Patients Benefit and at What Cost?
Authors: Pettinati Helen M. , Meyers Kathleen , Evans Bradley D. , Ruetsch Charles R. , Kaplan Frances N. , Jensen Jacqueline M. , Hadley Trevor R.
Published: July 2002

This study investigated whether selected patients have better outcomes with inpatient than outpatient treatment. There were 93 inpatients and 80 outpatients with alcohol dependence who were evaluated at treatment entry to a private healthcare setting. Patients with multiple drinking‐related consequences were less likely to return to significant drinking in the first 3 months after treatment ended if they had attended inpatient compared to outpatient treatment. Thus, inpatient appeared to have some advantage over outpatient treatment in the early recovery period for patients with multiple drinking‐related consequences. The gap between inpatient and outpatient costs was also reduced when computed as a cost‐effectiveness ratio, although treatment costs continued to remain proportionally higher with inpatient than outpatient treatment.
Introduction/Overview: Gender-Based Differences in Pharmacologic and Toxicologic Responses
Authors: Christian Mildred S.
Published: July 2002

Gender may be the most important factor in mammalian development and response to exogenous agents. From believing sex-related differences required sheltering women to protect their reproductive capacity (Victorians thought exercise, education, train travel, and certain music neuro-and reprotoxic to females) to legislating a status of essential equality of the sexes may have increased women's health issues. Men and women often respond differently to drugs. Inclusion of women in phase I/II clinical trials is insufficient to identify gender-based differences in response; rather, animal models should be the basis for predicting gender-based differences in pharmacologic and toxicologic effects. Unfortunately, current animal models do not consistently demonstrate such differences. Use of commonly used species (e.g., rats and dogs) does not necessarily result in relevant evaluation of an agent in a species at appropriate development (age), physiological state, anatomy, metabolism, or kinetics for estimation of human risks. The need to test agents in relevant animal models and advances in metabolic, pharmacokinetic, and pharmacodynamic capabilities challenge us to improve methods by using the most relevant models for estimating human risk. We need to be concerned about gender-related differences and the dynamics of gender-based growth and development over the entire life cycle. We must also consider potential interactions of dietary supplements and other exogenous agents that can act as drugs or modulate the potential effects of drugs differently in men, women, and developing children of both sexes. To this end, the health benefits of genistein and the effects of this dietary agent in a multigeneration study in rats will be described. It is envisioned that this symposium will assist in re-recognition of the importance of gender-related differences in use and response to Pharmaceuticals and result in optimization of nonclinical testing procedures to identify benefits and risks for human use of these agents.
On the Recognition of Multiple Chemical Sensitivity in Medical Literature and Government Policy
Authors: Donnay Albert H.
Published: July 2002

The history of chemical sensitivity in America is reviewed from the first description published by Edgar Allan Poe in 1839, to its first medical definition as a symptom of neurasthenia in 1869, its rediscovery as allergic toxemia in 1945, its redefinition in 1987 as multiple chemical sensitivity (MCS), and its overlap in the 1990s with chronic fatigue syndrome, fibromyalgia syndrome, and Gulf War syndrome (GWS). More than half of the over 500 peer-reviewed articles on MCS support an organic basis for MCS, whereas less than one-quarter support a psychiatric basis. The same 2:1 difference is seen in the numbers of MCS researchers writing these articles and the number of journals publishing them. A psychogenic interpretation of MCS also is specifically rejected in the latest formal position statement on the subject, a 1994 consensus of the American Lung Association, American Medical Association (AMA), U.S. Environmental Protection Agency (US EPA), and U.S. Consumer Product Safety Commission (US CPSC) (U.S. Government Printing Office 1994–523–217/81322). This and other government recognition of MCS in policy, research, and scientific conferences are summarized. Dozens of federal, state, and local authorities accept MCS as a legitimate disease and/or disability that deserves reasonable accommodation in housing, employment, and public facilities. Official recognition is expected later in 1999 when the U.S. Centers for Disease Control and Prevention (CDC) announces a formal definition of MCS and the federal Interagency Workgroup on MCS releases its long-awaited final report, 4 years in the making. Given that epidemiological data from three states puts the prevalence of chemical sensitivity at 16 to 33% of the general population, 2 to 6% of whom have already been diagnosed with MCS, this truly is a hidden epidemic that deserves the priority attention of public health researchers and policy makers. Industrial toxicologists are encouraged to work on reducing and eliminating the use of synthetic fragrances, chemical sensitizers, and other irritants in consumer products and occupational settings.
Profile of Patients with Chemical Injury and Sensitivity, Part II
Authors: Ziem Grace E.
Published: July 2002

Exposures which can induce multiple chemical sensitivity (MCS) involve symptomatic, usually repeated, exposures to pesticides, solvents, combustion products, remodeling, sick buildings, carbonless copy paper (occupational heavy use) and other irritants and petrochemicals. Accompanying toxic injury often involves the immune, endocrine and nervous systems as well as impairments in detoxification, energy and neurotransmitter metabolism, protein, mineral, and other nutrient deficiencies and gastrointestinal changes such as Candida, parasites, reduced chymotrypsin (marker enzyme for reduced pancreatic enzyme function), gluten intolerance, and reduced Secretory IgA. Chronic cortisol elevation leading to adrenal insufficiency if not corrected is common. Such elevation can lead to protein and mineral deficiencies with increased osteoporosis and reduced steroid precursors for normal estrogen and testosterone production. Detoxification changes often involve reduction in one or more Phase II pathways which causes excess free radical production. Impaired digestive enzymes can reduce breakdown of foods, with larger more antigenic molecules being absorbed and consequent food intolerances. Many of these conditions are treatable. There is extensive overlap of MCS with Chronic Fatigue Syndrome and Fibromyalgia which may be one condition in many cases. Current occupational exposure limits are not health based and thus may not prevent MCS and are totally inadequate to accommodate sensitive persons. Warning symptoms indicating increased risk for MCS onset include repeated headache, eye and respiratory irritation and fatigue. Eliminating exposures which cause repeated symptoms is a critical strategy for preventing sensitization and MCS. It also significantly reduces the degree of disability in persons with MCS, the single most important factor from the literature. Affected persons with disability can utilize the Americans With Disability Act to request reasonable accommodations for work, home (condo, apartment), and school.
Inhibitory Actions of Berberine on Growth and Arylamine N-Acetyltransferase Activity in Strains of Helicobacter Pylori from Peptic Ulcer Patients
Authors: Chung J. G. , Wu L. T. , Chang S. H. , Lo H. H. , Hsieh S. E. , Li Y. C. , Hung C. F.
Published: July 2002

N-acetyltransferase(NAT)-catalyzed reactions can result in activation or detoxification of arylamine carcinogens. Studies were conducted to examine the dose effects of berberine on inhibition of growth versus arylamine NAT activities in strains of Helicobacter pylori collected from peptic ulcer patients. An inhibition-of-growth study on H. pylori demonstrated that berberine elicited a dose-dependent growth inhibition in the H. Pylori cultures. The results on the NAT activity was determined using acetyl coenzyme A recycling assay and high pressure liquid chromatography. Cyto-sols or suspensions of H. pylori with or without selected concentrations of berberine cotreatment showed different percentages of 2-aminofluorene and p-aminobenzoic acid acetylation. The data indicated that there was decreased NAT activity associated with increased levels of berberine in the H. pylori cytosols and suspensions. The apparent values of Km and Vmax also decreased after cotreatment of berberine in both examined systems. This report is the first finding of berberine inhibition of arylamine NAT activity in strains of H. pylori.
Evaluation of an allelic association of the serotonin 5‐HT1B G681C polymorphism with antisocial alcoholism in the German population
Authors: Sander Thomas , Ostapowicz Agata , Samochowiec Jerzy , Smolka Michael , Rommelspacher Hans , Winterer Georg , Schmidt Lutz G.
Published: July 2002

Our study tested whether an association of the 861C allele of the serotonin 5‐HT1B gene (HTR1B) with antisocial alcoholism exists in the German population. The HTR1B G861C polymorphism was genotyped in 588 subjects of German descent, comprising 250 non‐alcoholic controls and 338 alcohol‐dependent subjects, of whom 56 exhibited a dissocial personality disorder (DSPD). The Tridimensional Personality Questionnaire was assessed in 109 alcohol‐dependent males to explore an effect of the 861C allele to risk‐taking behaviour. Our results revealed no evidence for an association of the 861C allele with antisocial alcoholism (p > 0.63). There were no significant differences in the personality traits, novelty‐seeking, harm avoidance and reward dependence between 46 male alcoholics carrying the 861C allele compared to those 63 alcoholics lacking it (p > 0.52). Our results do not provide evidence that the 861C allele contributes a substantial vulnerability effect to antisocial behavior in German alcohol‐dependent subjects.
Ascertainment of families for a linkage study of alcoholism
Authors: Cook Christopher C. H. , Hope Louise , Gurling Hugh
Published: July 2002

Traditionally, researchers working in the field of genetics and alcoholism have used treatment centres and clinics to try and recruit suitable subjects for research purposes. The current study considered a diverse range of possible sources to recruit suitable families for a linkage study of alcoholism. These sources included the press, personal contacts and circular letters to alcohol treatment centres and members of the Substance Misuse Section of the Royal College of Psychiatrists. Only 9–14% of families contacted from any source were suitable for inclusion in the study, due to the strict selection criteria. Press contacts were found to be the most productive source of suitable families willing to participate in the study, accounting for over 50% of contacts and eventual subjects recruited. There appeared to be no bias in the affection status of subjects recruited from the different sources. For future genetic studies of alcoholism it might be worthwhile to utilize this source more fully. Reasons for exclusion from the study are also considered, with the most common reasons being non‐co‐operation and no family history.
Hair testing for “ecstasy” (MDMA) in volunteer Scottish drug users
Authors: Ditton Jason , Cooper Gail A. A. , Scott Karen S. , Allen Desiree L. , Oliver John S. , Smith Iain D.
Published: July 2002

The aim of the study was to compare self reported “ecstasy” use with the results of the analysis of hair harvested from the same users. Subjects were recruited by multisite chain‐referral sampling within the 1994–95 “dance scene” in Glasgow. One hundred subjects donated hair after completing a lengthy interviewer‐administered questionnaire. Overall gross concordance between self reported “ecstasy” use and discovery of MDMA (or related compounds) in analysed hair did not surpass 59%, and no relationship had a Cohen's kappa of more than 0.08. Within the positive concordant dataset (n = 52), scatter was considerable, with no correlation being significant, and none more strongly positive than −0.0518. The results presented here indicate that, as far as MDMA is concerned, if judged by self‐report, hair does not reach a level of apparent accuracy that would permit its use as a general population estimator. However, hair testing is probably more reliable than self‐report, and its accuracy could be verified independently if large‐scale inter‐ and intra‐laboratory comparative research is conducted.
Parkinson's disease and aided AAC: some evidence from practice
Authors: Armstrong Linda , Jans Deborah , MacDonald Alison
Published: July 2002

Clinical observation that people with Parkinson's disease (PD) seem to have different training needs from other adult client‐groups in developing effective use of aided augmentative and alternative communication (AAC) was the catalyst for this study. There is little good‐quality research evidence available on the effectiveness (or lack of effectiveness) of aided AAC use by those with PD. One of the main aims of this preliminary study, therefore, was to elicit the experiences and attitudes of speech and language therapists in this specific area of their practice as the basis for future efficacy research and clinical practice. Thirty speech and language therapists were questioned about their experience of introducing low‐ and high‐tech AAC devices to this client‐group. Of particular interest was the discovery of the factors identified by the therapists as influencing their introduction of aided AAC to someone with PD and affecting implementation and successful use. The factors reported were both disease‐specific (e.g. motor problems) and more general factors (e.g. carer support) relevant to the process of introducing aided AAC to other adult client‐groups. Implications for future research and practice are discussed. Some research questions are posed and the inclusion of people with PD and their carers in practice development is promoted.
Essential Role of the Mitochondrial Respiratory Chain in Peroxynitrite‐Induced Strand Scission of Genomic DNA
Authors: Guidarelli Andrea , Tommasini Ilaria , Fiorani Mara , Cantoni Orazio
Published: July 2002

A large body of experimental evidence suggests that DNA damage and cytotoxicity mediated by peroxynitrite are linked by a causal relationship and important events in various pathological conditions. In the present study, we investigated the mechanism whereby peroxynitrite causes DNA single strand breakage in intact cells and found that the respiratory chain plays a pivotal role in this response. In particular, peroxynitrite mediates inhibition of complex III and, under these conditions, electrons are directly transferred from ubisemiquinone to molecular oxygen. Hydrogen peroxide produced by the dismutation of superoxides is the species mediating the peroxynitrite‐dependent DNA cleavage.
Quantitative Determination of Surface Concentration of Human Apolipoprotein H with Capillary Electrophoresis
Authors: Wang Shao‐xiong , Sun Yu‐Tong , Ru Qin‐hua , Luo Guo‐an , Sui Sen‐fang
Published: July 2002

The phospholipid monolayer at an air/water interface is widely used to mimic the biological membrane. The dynamic process of the protein or peptide interacting with lipid molecules can be reflected in the change in surface pressure of the monolayer. But the conventional method used to measure the surface pressure change gives results that cannot easily be correlated with the contribution of a single protein molecule. Previously, measuring the surface concentration of the protein molecules at the air/water interface has required the protein to be labeled with radioactivity or fluorescence. Here, a new method using capillary electrophoresis is introduced to measure the surface concentration of the protein. The results show at least two advantages of the new method: The numerical results of protein concentration can be obtained in a more precise and rapid way; and there is no need to label the protein sample or to build a special monolayer setup.
Effect of Buffer Solutions on Activation of Shamouti Orange Pyrophosphate‐Dependent Phosphofructokinase by Fructose 2,6‐Bisphosphate
Authors: Van Praag Esther , Tzur Assaf , Zehavi Uri , Goren Raphael
Published: July 2002

Shamouti phosphofructokinase (PFP) activation depends on the presence of fructose 2,6‐bisphosphate (Fru‐2,6‐P2) in the glycolytic reaction. The effect of activation by Fru‐2,6‐P2 differs considerably, however, according to the buffer (pH 8.0) in which the reaction is performed: Ka = 2.77 plus/minus 0.3 nM in Hepes‐NaOH and 7.75 plus/minus 1.49 nM in Tris‐HCl. The presence of chloride ions (39 mM) in the Tris‐HCl buffer inhibits PFP. Indeed, when using a Hepes‐NaOH buffer and then adding 39 mM NaCl, Ka = 8.12 plus/minus 0.52 nM. The Ki for chloride ions is approximately 21.7 mM. In the gluconeogenic reaction, Shamouti PFP generally showed a high endogenous activity. Addition of Fru‐2,6‐P2 did not modify the velocity and the Vmax of the enzyme; however, its presence increased the affinity of the enzyme for Fru‐1,6‐P2 from 200 plus/minus 15.6 muM in absence of Fru‐2,6‐P2 to 89 plus/minus 10.3 muM in its presence (10 muM). In the presence of chloride (39 mM), the affinity for the substrate decreased with Km = 150 plus/minus 14 muM. The calculated Ki for chloride ions equals 56.9 mM. In both the glycolytic and the gluconeogenic reactions, Vmax is not affected; therefore, the inhibition mode of chloride is competitive.
Selection of Peptide Ligands That Bind to Acid Fibroblast Growth Factor
Authors: Fan Hongkuan , Liu Ying , Zhou Hui , Wang Liping , Li Wei , Guo Lili , Roeske Roger W.
Published: July 2002

Acid fibroblast growth factor (aFGF) binds to its cell‐surface receptors in a heparin‐dependent manner. In an attempt to define the aFGF recognition site on fibroblast growth factor receptor 1 (FGFR1), we developed a screening strategy for identifying FGF ligands that bind to the receptor‐binding region of FGF. To retain the natural conformation of aFGF during screening, we used biotinylated heparin to immobilize aFGF on a streptavidincoated dish. A 15‐mer phage display peptide library was then screened in the dish and a group of related peptide sequences was identified. These peptide sequences contain two conserved motifs, SSG and VPS, corresponding to two protein sequences of the immunoglobulin‐like (Ig‐like) domain II of FGFR1 at amino acids 180‐182 and 221‐223 (CPSSG‐VPSDKGNYTC). Further experiments demonstrate that the phage displaying these sequences can specifically bind to aFGF and that the synthesized peptide corresponding in sequence can inhibit mitogenic activity of aFGF.These sequences may thus constitute part of the aFGF‐binding region on FGFR1, and the synthesized peptide has the potential to become a therapeutic agent as an aFGF antagonist.
Latent Transforming Growth Factor-β Binding Protein Domains Involved in Activation and Transglutaminase-dependent Cross-Linking of Latent Transforming Growth Factor-β
Authors: Nunes Irene , Gleizes Pierre-Emmanuel , Metz Christine N. , Rifkin Daniel B
Published: July 2002

Transforming growth factor-β (TGF-β) is secreted by many cell types as part of a large latent complex composed of three subunits: TGF-β, the TGF-β propeptide, and the latent TGF-β binding protein (LTBP). To interact with its cell surface receptors, TGF-β must be released from the latent complex by disrupting noncovalent interactions between mature TGF-β and its propeptide. Previously, we identified LTBP-1 and transglutaminase, a cross-linking enzyme, as reactants involved in the formation of TGF-β. In this study, we demonstrate that LTBP-1 and large latent complex are substrates for transglutaminase. Furthermore, we show that the covalent association between LTBP-1 and the extracellular matrix is transglutaminase dependent, as little LTBP-1 is recovered from matrix digests prepared from cultures treated with transglutaminase inhibitors. Three polyclonal antisera to glutathione S–transferase fusion proteins containing amino, middle, or carboxyl regions of LTBP-1S were used to identify domains of LTBP-1 involved in crosslinking and formation of TGF-β by transglutaminase. Antibodies to the amino and carboxyl regions of LTBP-1S abrogate TGF-β generation by vascular cell cocultures or macrophages. However, only antibodies to the amino-terminal region of LTBP-1 block transglutaminase-dependent cross-linking of large latent complex or LTBP-1. To further identify transglutaminase-reactive domains within the amino-terminal region of LTBP-1S, mutants of LTBP-1S with deletions of either the amino-terminal 293 (ΔN293) or 441 (ΔN441) amino acids were expressed transiently in CHO cells. Analysis of the LTBP-1S content in matrices of transfected CHO cultures revealed that ΔN293 LTBP-1S was matrix associated via a transglutaminasedependent reaction, whereas ΔN441 LTBP-1S was not. This suggests that residues 294–441 are critical to the transglutaminase reactivity of LTBP-1S.
The Isolated Comet Tail Pseudopodium of Listeria monocytogenes: A Tail of Two Actin Filament Populations, Long and Axial and Short and Random
Authors: Sechi Antonio S. , Wehland Jürgen , Small J. Victor
Published: July 2002

Listeria monocytogenes is driven through infected host cytoplasm by a comet tail of actin filaments that serves to project the bacterium out of the cell surface, in pseudopodia, to invade neighboring cells. The characteristics of pseudopodia differ according to the infected cell type. In PtK2 cells, they reach a maximum length of ∼15 μm and can gyrate actively for several minutes before reentering the same or an adjacent cell. In contrast, the pseudopodia of the macrophage cell line DMBM5 can extend to >100 μm in length, with the bacteria at their tips moving at the same speed as when at the head of comet tails in bulk cytoplasm. We have now isolated the pseudopodia from PtK2 cells and macrophages and determined the organization of actin filaments within them. It is shown that they possess a major component of long actin filaments that are more or less splayed out in the region proximal to the bacterium and form a bundle along the remainder of the tail. This axial component of filaments is traversed by variable numbers of short, randomly arranged filaments whose number decays along the length of the pseudopodium. The tapering of the tail is attributed to a grading in length of the long, axial filaments.

The exit of a comet tail from bulk cytoplasm into a pseudopodium is associated with a reduction in total F-actin, as judged by phalloidin staining, the shedding of α-actinin, and the accumulation of ezrin. We propose that this transition reflects the loss of a major complement of short, random filaments from the comet, and that these filaments are mainly required to maintain the bundled form of the tail when its borders are not restrained by an enveloping pseudopodium membrane. A simple model is put forward to explain the origin of the axial and randomly oriented filaments in the comet tail.
mRNAs for Microtubule Proteins Are Specifically Colocalized during the Sequential Formation of Basal Body, Flagella, and Cytoskeletal Microtubules in the Differentiation of Naegleria gruberi
Authors: Han Ji Woong , Park Jong Ho , Kim Misook , Lee JooHun
Published: July 2002

We have examined the distribution of four mRNAs—α-tubulin, β-tubulin, flagellar calmodulin, and Class I mRNA—during differentiation of Naegleria gruberi amebas into flagellates by in situ hybridization. Three of the four mRNAs—α-tubulin, β-tubulin, and Class I mRNA—began to be colocalized at the periphery of the cells as soon as transcription of the respective genes was activated and before any microtubular structures were observable. At 70 min after the initiation of differentiation, these mRNAs were relocalized to the base of the growing flagella, adjacent to the basal bodies and microtubule organizing center for the cytoskeletal microtubules. Within an additional 15 min, the mRNAs were translocated to the posterior of the flagellated cells, and by the end of differentiation (120 min), very low levels of the mRNAs were observed. Cytochalasin D inhibited stage-specific localization of the mRNAs, demonstrating that RNA localization was actin dependent. Since cytochalasin D also blocked differentiation, this raises the possibility that actin-dependent RNA movement is an essential process for differentiation.
Regulated Production of Mineralization-competent Matrix Vesicles in Hypertrophic Chondrocytes
Authors: Kirsch Thorsten , Nah Hyun-Duck , Shapiro Irving M. , Pacifici Maurizio
Published: July 2002

Matrix vesicles have a critical role in the initiation of mineral deposition in skeletal tissues, but the ways in which they exert this key function remain poorly understood. This issue is made even more intriguing by the fact that matrix vesicles are also present in nonmineralizing tissues. Thus, we tested the novel hypothesis that matrix vesicles produced and released by mineralizing cells are structurally and functionally different from those released by nonmineralizing cells. To test this hypothesis, we made use of cultures of chick embryonic hypertrophic chondrocytes in which mineralization was triggered by treatment with vitamin C and phosphate. Ultrastructural analysis revealed that both control nonmineralizing and vitamin C/phosphatetreated mineralizing chondrocytes produced and released matrix vesicles that exhibited similar round shape, smooth contour, and average size. However, unlike control vesicles, those produced by mineralizing chondrocytes had very strong alkaline phosphatase activity and contained annexin V, a membrane-associated protein known to mediate Ca2+ influx into matrix vesicles. Strikingly, these vesicles also formed numerous apatite-like crystals upon incubation with synthetic cartilage lymph, while control vesicles failed to do so. Northern blot and immunohistochemical analyses showed that the production and release of annexin V-rich matrix vesicles by mineralizing chondrocytes were accompanied by a marked increase in annexin V expression and, interestingly, were followed by increased expression of type I collagen. Studies on embryonic cartilages demonstrated a similar sequence of phenotypic changes during the mineralization process in vivo. Thus, chondrocytes located in the hypertrophic zone of chick embryo tibial growth plate were characterized by strong annexin V expression, and those located at the chondro–osseous mineralizing border exhibited expression of both annexin V and type I collagen. These findings reveal that hypertrophic chondrocytes can qualitatively modulate their production of matrix vesicles and only when induced to initiate mineralization, will release mineralization-competent matrix vesicles rich in annexin V and alkaline phosphatase. The occurrence of type I collagen in concert with cartilage matrix calcification suggests that the protein may facilitate crystal growth after rupture of the matrix vesicle membrane; it may also offer a smooth transition from mineralized type II/type X collagen-rich cartilage matrix to type I collagen-rich bone matrix.
Laminin 5 Binds the NC-1 Domain of Type VII Collagen
Authors: Rousselle Patricia , Keene Douglas R. , Ruggiero Florence , Champliaud Marie-France , Rest Michel van der , Burgeson Robert E.
Published: July 2002

Mutational analyses of genes that encode components of the anchoring complex underlying the basolateral surface of external epithelia indicate that this structure is the major element providing for resistance to external friction. Ultrastructurally, laminin 5 (α3β3γ2; a component of the anchoring filament) appears as a thin filament bridging the hemidesmosome with the anchoring fibrils. Laminin 5 binds the cell surface through hemidesmosomal integrin α6β4. However, the interaction of laminin 5 with the anchoring fibril (type VII collagen) has not been elucidated. In this study we demonstrate that monomeric laminin 5 binds the NH2-terminal NC-1 domain of type VII collagen. The binding is dependent upon the native conformation of both laminin 5 and type VII collagen NC-1. Laminin 6 (α3β1γ1) has no detectable affinity for type VII collagen NC-1, indicating that the binding is mediated by the β3 and/or γ2 chains of laminin 5. Approximately half of the laminin 5 solubilized from human amnion or skin is covalently complexed with laminins 6 or 7 (α3β2γ1). The adduction occurs between the NH2 terminus of laminin 5 and the branch point of the short arms of laminins 6 or 7. The results are consistent with the presumed orientation of laminin 5, having the COOH-terminal G domain apposed to the hemidesmosomal integrin, and the NH2-terminal domains within the lamina densa. The results also support a model predicting that monomeric laminin 5 constitutes the anchoring filaments and bridges integrin α6β4 with type VII collagen, and the laminin 5–6/7 complexes are present within the interhemidesmosomal spaces bound at least by integrin α3β1 where they may mediate basement membrane assembly or stability, but contribute less significantly to epithelial friction resistance.
Myelin-associated Glycoprotein Interacts with Neurons via a Sialic Acid Binding Site at ARG118 and a Distinct Neurite Inhibition Site
Authors: Tang Song , Shen Ying Jing , DeBellard Maria Elena , Mukhopadhyay Gitali , Salzer James L. , Crocker Paul R. , Filbin Marie T.
Published: July 2002

Inhibitory components in myelin are largely responsible for the lack of regeneration in the mammalian CNS. Myelin-associated glycoprotein (MAG), a sialic acid binding protein and a component of myelin, is a potent inhibitor of neurite outgrowth from a variety of neurons both in vitro and in vivo. Here, we show that MAG's sialic acid binding site is distinct from its neurite inhibitory activity. Alone, sialic acid–dependent binding of MAG to neurons is insufficient to effect inhibition of axonal growth. Thus, while soluble MAG-Fc (MAG extracellular domain fused to Fc), a truncated form of MAG-Fc missing Ig-domains 4 and 5, MAG(d1-3)-Fc, and another sialic acid binding protein, sialoadhesin, each bind to neurons in a sialic acid– dependent manner, only full-length MAG-Fc inhibits neurite outgrowth. These results suggest that a second site must exist on MAG which elicits this response. Consistent with this model, mutation of arginine 118 (R118) in MAG to either alanine or aspartate abolishes its sialic acid–dependent binding. However, when expressed at the surface of either CHO or Schwann cells, R118-mutated MAG retains the ability to inhibit axonal outgrowth. Hence, MAG has two recognition sites for neurons, the sialic acid binding site at R118 and a distinct inhibition site which is absent from the first three Ig domains.
Nef-mediated Clathrin-coated Pit Formation
Authors: Foti Michelangelo , Mangasarian Aram , Piguet Vincent , Lew Daniel P. , Krause Karl-Heinz , Trono Didier , Carpentier Jean-Louis
Published: July 2002

The sequence of events leading to clathrin-coated pit (CCP) nucleation on the cell surface and to the incorporation of receptors into these endocytic structures is still imperfectly understood. In particular, the question remains as to whether receptor tails initiate the assembly of the coat proteins or whether receptors migrate into preformed CCP. This question was approached through a dissection of the mechanisms implemented by Nef, an early protein of human and simian immunodeficiency virus (HIV and SIV, respectively), to accelerate the endocytosis of cluster of differentiation antigen type 4 (CD4), the major receptor for these viruses. Results collected showed that: (a) Nef promotes CD4 internalization via an increased association of CD4 with CCP; (b) the Nef-mediated increase of CD4 association with CCP is related to a doubling of the plasma membrane area occupied by clathrin-coated structures; (c) this increased CCP number at the plasma membrane has functional consequences preferentially on CD4 uptake and does not significantly affect transferrin receptor internalization or fluid-phase endocytosis; (d) the presence of a CD4 cytoplasmic tail including a critical dileucine motif is required to induce CCP formation via Nef; and (e) when directly anchored to the cytoplasmic side of the plasma membrane, Nef itself can promote CCP formation. Taken together, these observations lead us to propose that CD4 can promote CCP generation via the connector molecule Nef. In this model, Nef interacts on one side with CD4 through a dileucine-based motif present on CD4 cytoplasmic tail and on the other side with components of clathrin-coated surface domain (i.e., adaptins). These Nef-generated complexes would then initiate the nucleation of CCP.
Phosphatidylinositol 3–Kinase Is Required for the Formation of Constitutive Transport Vesicles from the TGN
Authors: Jones Steven M. , Howell Kathryn E.
Published: July 2002

An 85-kD cytosolic complex (p62cplx), consisting of a 62-kD phosphoprotein (p62) and a 25-kD GTPase, has been shown to be essential for the cell-free reconstitution of polymeric IgA receptor (pIgA-R)-containing exocytic transport vesicle formation from the TGN (Jones, S.M., J.R. Crosby, J. Salamero, and K.E. Howell. 1993. J. Cell Biol. 122:775–788). Here the p62cplx is identified as a regulatory subunit of a novel phosphatidylinositol 3–kinase (PI3-kinase). This p62cplx-associated PI3-kinase activity is stimulated by activation of the p62cplx-associated GTPase, and is specific for phosphatidylinositol (PI) as substrate, and is sensitive to wortmannin at micromolar concentrations. The direct role of this p62cplx-associated PI3-kinase activity in TGN-derived vesicle formation is indicated by the finding that both lipid kinase activity and the formation of pIgA-R–containing exocytic vesicles from the TGN are inhibited by wortmannin with similar dose-response curves and 50% inhibitory concentrations (3.5 μM). These findings indicate that phosphatidylinositol-3-phosphate (PI[3]P) is required for the formation of TGN-derived exocytic transport vesicles, and that the p62cplx-associated PI3-kinase and an activated GTPase are the essential molecules that drive production of this PI(3)P.
The Ras Target AF-6 Interacts with ZO-1 and Serves as a Peripheral Component of Tight Junctions in Epithelial Cells
Authors: Yamamoto Takaharu , Harada Naozumi , Kano Kyoko , Taya Shin-ichiro , Canaani Eli , Matsuura Yoshiharu , Mizoguchi Akira , Ide Chizuka , Kaibuchi Kozo
Published: July 2002

The dynamic rearrangement of cell–cell junctions such as tight junctions and adherens junctions is a critical step in various cellular processes, including establishment of epithelial cell polarity and developmental patterning. Tight junctions are mediated by molecules such as occludin and its associated ZO-1 and ZO-2, and adherens junctions are mediated by adhesion molecules such as cadherin and its associated catenins. The transformation of epithelial cells by activated Ras results in the perturbation of cell–cell contacts. We previously identified the ALL-1 fusion partner from chromosome 6 (AF-6) as a Ras target. AF-6 has the PDZ domain, which is thought to localize AF-6 at the specialized sites of plasma membranes such as cell–cell contact sites. We investigated roles of Ras and AF-6 in the regulation of cell–cell contacts and found that AF-6 accumulated at the cell–cell contact sites of polarized MDCKII epithelial cells and had a distribution similar to that of ZO-1 but somewhat different from those of catenins. Immunoelectron microscopy revealed a close association between AF-6 and ZO-1 at the tight junctions of MDCKII cells. Native and recombinant AF-6 interacted with ZO-1 in vitro. ZO-1 interacted with the Ras-binding domain of AF-6, and this interaction was inhibited by activated Ras. AF-6 accumulated with ZO-1 at the cell–cell contact sites in cells lacking tight junctions such as Rat1 fibroblasts and PC12 rat pheochromocytoma cells. The overexpression of activated Ras in Rat1 cells resulted in the perturbation of cell–cell contacts, followed by a decrease of the accumulation of AF-6 and ZO-1 at the cell surface. These results indicate that AF-6 serves as one of the peripheral components of tight junctions in epithelial cells and cell–cell adhesions in nonepithelial cells, and that AF-6 may participate in the regulation of cell–cell contacts, including tight junctions, via direct interaction with ZO-1 downstream of Ras.