bluuebunny/crossref_metadata_2025 huggingface dataset:
Changing Employment Systems in China: Some Evidence from the Shenzhen Special Economic Zone
Authors: Ip Olivia K.M.
Published: June 2007

A stated objective of the reform of employment systems promulgated by the government in China has been to instil some `free labour market mechanisms' in contrast to the pre-existing highly centralised administered `allocation systems'. This paper seeks to examine the changing patterns of employment against the background of labour reforms in China since 1979. Drawing on case studies conducted in five enterprises in the Shenzhen Special Economic Zone, it is argued that, despite the evident phenomenal geographical mobility of labour, it appears that the labour force remains very stratified. It is further argued that, paradoxically, the government's institutional barriers appear to be a major cause of such stratification.
Labour Regimes and the Domestic Domain: Manufacturing Garments in Rural Greece
Authors: Simmons Colin , Kalantaridis Christos
Published: June 2007

Over the last quarter of a century, Greece, alongside other southern European economies, witnessed a burgeoning of new enterprises and the creation of a significant number of jobs in the garment industry. Many producers were beneficiaries of the decentralization of production from large corporations in core countries (particularly Germany) to localised networks, through sub-contracting arrangements. Our paper highlights the case of garment-led industrial growth in the Macedonian countryside. We set out to explore the nature of the labour regime which lies at the heart of the recent expansion. The argument is that medium-scale primary sub-contractors have introduced novel practices of employment in the locality, while their household and small-scale secondary counterparts have based their enterprise strategy upon redefining their links with traditional institutions, especially the family-oriented domestic domain.
Automation and New Work Patterns: Cases from Singapore's Electronics Industry
Authors: Yun Hing Ai
Published: June 2007

In Singapore, the state has a policy of the extensive encouragement of automation as well as the continuous upgrading of the labour force through further education and retraining. This paper reports on a survey of workers, together with case studies of firms, in the electronics industry. It suggests that the implementation of automation may at times be in conflict with the training and educational upgrading policy, and may be accompanied by extensive labour turnover due to the demands of new working hours associated with automation. However, the high level of demand for labour within Singapore, and the ready availability of alternative sources of supply from outside, means that this is not a serious problem. In general, workers considered that their levels of skill had been enhanced as a result of automation, but also that the intensity of their effort had been increased.
Employees' Perspectives on One Dimension of Labour Flexibility: Working at a Distance
Authors: Hamblin Heather
Published: June 2007

This article argues that working at a distance (that is, work done in the home at a physical distance from the employer, using information-based electronic equipment for some parts of the job) will become an increasingly significant phenomenon towards the twenty-first century. This development can bring benefits to both the employer and the individual, although other concerns regarding remote work for the individual and the organisation are discussed fully. Since the literature to date has considered working at a distance mainly from the employer's perspective, and largely in respect of professional and computing staffs, the article presents evidence from some empirical research undertaken as part of a larger study into labour flexibility: the analysis of secretarial employees' attitudes towards the flexibility of their work location and possible teleworking from home. The results, including the preferred option of working part of the week at home, are reviewed and their potential importance set within the wider labour flexibility and human resource management agendas.
Self-Employment Career Dynamics: The Case of `Unemployment Push' in UK Book Publishing
Authors: Granger Bill , Stanworth John , Stanworth Celia
Published: June 2007

The recent revival of self-employment in the UK and other advanced industrialized economies has been viewed contrastingly as an indication of economic vitality and, alternatively, as a form of labour market deficiency. These different perceptions rest essentially on two opposing processes of entry into self-employment -'entrepreneurial pull' and `unemployment push'. The research reported here, into freelancing in book publishing, reveals patterns of entry into self-employment which reflect the presence of both these processes, plus additional configurations and changes over time. The respondents, being predominantly female, were ultra-typical of those who swelled the self-employed workforce during the 1980s, when the number of female self-employed without employees doubled in the period 1981-93.
The Relationship between Working Life and Individualisation: A Study among Danish Trade Union Members
Authors: Madsen Morten
Published: June 2007

A significant current trend in the industrialised countries is a growing individualization among wages-earners: i.e changes from collective value orientations based on solidarity and equality towards more individualistic value orientations based on self-interest and personal opportunities. This paper analyses relations between these changes in wage-earners' identity, labour market position, job characteristics, and relationship to the company and trade union: both empirically, by using Danish survey data from the research project: `The Employee Perspective on Working Life and Politics', and theoretically, by discussing the contrasting ideas of Hirsch and Roth (1986) and Valkenburg (1995) and Zoll (1995). The analysis shows both expected and surprising relationships between labour market positions, job characteristics and individualization. Furthermore, the paper demonstrates that the term individualization' must be understood as a relative concept, and that an increased individualization does not necessarily lead towards a dissolution of the trade union movement as a representative of wage-earners' interests in modern society.
Developments in Trade Unionism in the Financial Sector in Britain
Authors: Gall Gregor
Published: June 2007

This article puts forward evidence to argue that finance workers, especially bank workers, in Britain are becoming more `unionate'. This is not to argue that these workers are becoming `militant' for while the developments are significant they remain cautious and uneven. An examination is made of why these developments have occurred and how they relate to the employers' human resource policies, why the growth in trade unionism in the financial sector runs counter to a trend of decline elsewhere and why significant differences in trade unionism exist within the different components in the financial sector.
The Managerial Career after Downsizing: Case Studies from the `Leading Edge'
Authors: McGovern Patrick , Hope-Hailey Veronica , Stiles Philip
Published: June 2007

This paper sets out to investigate whether the recent wave of organisational restructuring has contributed to the further decline of the internally promoted manager or produced a new model of managerial employment in large organizations. Our research, which is based on in-depth case studies of eight major British-based employers, finds no evidence of the kind of transformational change associated with the introduction of a new model. Instead, we find that the traditional model of managerial employment has been eroded rather than replaced. The most notable changes include less job security - especially for those older than fifty - more emphasis on `managing your own career', fewer opportunities for upward promotion, and an increased emphasis on lateral career moves. We conclude by arguing that this restructuring adds further impetus to the decline of the internally promoted middle manager.
Caring for Everyone? The Implications for Women of the Changes in Community Care Services
Authors: Smith Helen
Published: June 2007

In Britain, services for people with all forms of disabilities have been undergoing major structural change. However, despite the stake that women have in community care services, as users, carers and workers, a feminist viewpoint has not featured at all in the debate. The stakes are high for women, and feminists need to understand the import of these changes when seen within a gender-aware political context. This article outlines the changes that are occurring in our community care services and draws out the implications for women from a feminist perspective.
Mothers on Trial: The Custodial Vulnerability of Women
Authors: Chesler Phyllis
Published: June 2007

Mothers are not protected by patriarchal law or custom. Double standards pervade the child custody courts: fathering is over-valued, mothering under-valued. Fathers often win custody even when they are parentally uninvolved or abusive; mothers lose it for any departure from an idealized stereotype of motherhood. In particular, male violence is routinely minimized and female response to it seen as pathological. Creative tactics are necessary in order for custodially embattled mothers to survive; and more publicity and more facilities are urgently needed.
Feminism and Discourse in Developmental Psychology: Power, Subjectivity and Interpretation
Authors: Burman Erica
Published: June 2007

The meeting of feminism with post-structuralism has brought issues of multiplicity of meanings to the fore. However, attention to difference threatens to disperse politics. This paper explores how these issues are played out in developmental psychological research. The concerns of feminist research with issues of reciprocity, consultation and accountability challenge developmental research conducted within the structural power relations of adult-child, experimenter-subject encounters. Equally, the feminist project of social transformation both draws upon and renders problematic developmental psychology. Drawing on examples from my interviews with children, I argue that attending to the power relations that are conferred by structural research practices and subject positions set up within discourse not only highlights previously unacknowledged diversity of meaning, but also closes off total interpretive relativism. What we need is a feminist realism that does not resort to positivism, that ties discourse to politics, and that makes politics more than discursive.
Sex, Organs and Audiotape: A Discourse Analytic Approach to Talking about Heterosexual Sex and Relationships
Authors: Gilfoyle Jackie , Wilson Jonathan , Own Br
Published: June 2007

This article examines talk about sex and heterosexual relationships, based on a study of 12 women and 13 men who participated in semi-structured interviews, in order to identify the `discourses' of sexuality which inform talk about heterosexual sex. One theme in talk about heterosexuality can be understood through the `pseudo-reciprocal gift discourse': women are described as `giving' themselves to men, whereas men `give' women orgasms, reproducing dominant norms of male activity and female passivity, — and thereby reinforcing the oppression of women. Men talk more graphically about sex than women — we suggest the resources of meaning concerning sex suit men's interests rather than women's, and reflect men's dominance in a (hetero)sexist society.
`Wild Women Don't Have the Blues': A Feminist Critique of `Person-Centred' Counselling and Therapy
Authors: Waterhouse Ruth L.
Published: June 2007

Contemporary feminists have been concerned to address the impact of psychoanalysis on the social construction of femininity. By contrast, the work of humanistic psychologists has received relatively little in the way of feminist scrutiny. This is apparent within person-centred counselling which can be regarded as a rapidly developing social movement in contemporary Britain. The person-centred approach has been adopted not only by `mainstream' agencies but by many feminist agencies and women `s self-help groups. Drawing on the insights of feminist philosophy this article attempts to subject the key concepts of Rogerian counselling to gender analysis. The so-called `core conditions' are addressed from a feminist perspective and the principles of `responsibility' and `authenticity' examined from a woman-centred viewpoint. The problems of adopting Rogerian principles in the context of Rape Crisis counselling are addressed. A consideration is made of the extent to which humanism and the counselling practices informed by this tradition have failed to address the specific problems encountered by women in their search for personhood and agency.
Giving Evidence: Women's Lives in Ashworth Maximum Security Psychiatric Hospital
Authors: Potier Moira A.
Published: June 2007

Based on written evidence given to the Public Inquiry on Ashworth Maximum Security Psychiatric Hospital in Britain, this article graphically documents the conditions of women patients' lives at Ashworth. It exposes systematic abuses of power by hospital staff, and the techniques used to suppress women's resistance to such abuse. A postscript reports on the situation at Ashworth today, in the face of attempts made to implement the recommendations of the Commission of Inquiry.
Remaking the Link: Qualitative Research and Feminist Standpoint Theory
Authors: Henwood Karen , Pidgeon Nick
Published: June 2007

Historical and contemporary developments in feminist thinking on theory and method are considered. The critique of positivism is outlined, together with the affinity between feminist research and qualitative methods. Dilemmas are raised concerning: a false dualism between quantity and quality; ethical concerns; and writing in the personal. There are contradictory positions within the contemporary argument for methodological pluralism, such that it does not resolve tensions surrounding issues of women's experience and the warranting of interpretations. Epistemological debates involving feminism and post-modernism have created a changed intellectual landscape for reconsidering the link between qualitative research and standpoint theory. These raise issues of grounding, depth and evaluating knowledge by appeals to a notion of strong (or feminist) objectivity, together with the need to mitigate against appropriation of the `other'. Exemplars of qualitative feminist analysis are presented.
`Race', Racism, and Sexuality in the Life Narratives of Immigrant Women
Authors: Espin Oliva M.
Published: June 2007

Life narratives were collected from immigrant women through five individual interviews and three focus groups as part of an on-going study of sexuality and gender-related issues among immigrant/refugee women in North America. The research explores how questions of national identity and sexual identity are determined and negotiated for immigrant and refugee women and the role of `geography' and language in understanding women's boundary and border crossings. Four life stories are discussed in detail, as illustrative of disparate adaptations to migration, and suggestions are made for the future development of research in this area.
Dealing with Difficult Differences: Reflexivity and Social Class in Feminist Research
Authors: Reay Diane
Published: June 2007

This article explores the impact of difference within the research process. First, it looks at the difference my working-class history has made to the experience of conducting feminist research. Second, it examines how that class difference has affected both my relationships with the women that I inter-viewed and my reading of their accounts. Drawing on both my own and the women s stories of growing up working class, I have attempted to map out some of the psychological complexities that are invariably overlooked in academic accounts of either-the research process or working-class experiences of mother-ing and childhood. Finally, I have tried to pull out some of the threads still linking educated, working-class, feminist academics with their class backgrounds and explore the fragility and strength of those connections in the light of my own social trajectory.
`Wham Bam, am I a Man?': Unemployed Men Talk about Masculinities
Authors: Willott Sara , Griffin Christine
Published: June 2007

In contemporary society, being powerful is typically associated with, among other things, being male, middle class and employed. The cultural ascendancy of these characteristics is supported by specific structural and discursive patterns. However, there are a number of ways in which these cultural yardsticks can be challenged. In this paper we summarize the discursive patterns constructed by a group of working-class men experiencing long-term unemployment in a region of the English West Midlands. These men talked about a conflict between discourses concerning domestic provision and public consumption, leading to a sense of disempowerment and emasculation. Despite the potential challenge posed by long-term unemployment to traditional versions of masculinity, these men's accounts retained their positions within hegemonic discourses of masculinity. Finally, we examine the political implications of such discursive patterns.
Body Image Perception among Women of African Descent: A Normative Context?
Authors: Ofosu Helen B. , Lafreniere Kathryn D. , Senn Charlene Y.
Published: June 2007

The purpose of this article is to integrate research from across the social sciences in order to understand Black women's body image perceptions. To accomplish this goal, the effects of race, ethnicity, class and culture are examined. Cultural differences between African Canadians/Americans and Blacks living in Africa and the Caribbean are outlined and their effects on Black women's experiences of weight and eating are also discussed. Some of the historical underpinnings, and several aspects of the current social context within which Black women live are explored in order to explain body image perceptions.
Women Construct Self-Esteem in their Own Terms: A Feminist Qualitative Study
Authors: Chatham-Carpenter April , DeFrancisco Victoria
Published: June 2007

The purpose of this article is to explore the concept of self-esteem from adult women's perspectives. Based on qualitative interviews with 59 women, we found similar denotative definitions of self-esteem, but differing connotative views and experiences. The women identified three overall characteristics associated with high self-esteem (i.e. `voice', `self-perspective' and `concern for others'), viewing them as interconnected rather than isolated variables. We found self-esteem to be a reflexive, context-specific process, which is changeable over the lifespan. The women's constructions of self-esteem as a complex phenomenon are in contrast to traditional androcentric and essentialist self-esteem scales in common usage. Their views of self-esteem were socially constructed in dialogue with their culture, other persons and themselves. Implications of taking such a social constructivist approach to studying self-esteem are discussed.
Recovering a Woman's Head With Prophetic Authority: a Pentecostal Interpretation of 1 Corinthians 11.3-16
Authors: Powers Janet
Published: June 2007

Early Pentecostals defended the ministry of women by using passages in Acts 2.16-17 and 1 Cor. 11.3-16 to show that the Holy Spirit had empowered women to prophesy. But in Pentecostal churches today, some of the same biblical passages are now used to argue for significant restrictions on the ministry of women. This shift is especially apparent in the interpretation of 1 Cor. 11.3-16.These contemporary Pentecostals do not seem to realize that the hermeneutic that is used to interpret 1 Cor. 11.3-16 as a passage which limits the ministry of women is the same hermeneutic which is often used to discredit the doctrine of Spirit-baptism. Pentecostal interpreters need to reject this non-Pentecostal hermeneutic and reclaim 1 Cor. 11.3-16 as part of the Pentecostal defense of Spirit-empowered ministries of women. What is at stake is not just the prophetic ministry of women but the fundamental Pentecostal belief that all believers are empowered by the Holy Spirit for ministry.
Stretching the Definitions? Pneumatology and 'Syncretism' in African Pentecostalism
Authors: Anderson Allan
Published: June 2007

Some observers regard as 'syncretistic' many forms of African Pentecostalism that have developed a pneumatology with a presumed link to the pre-Christian past, and will also deny the term 'Pentecostal' to these groups. This does not fully recognise the parallels between biblical pneumatology and the holistic African worldview, and the significant contribution that African Pentecostalism makes to a dynamic understanding of pneumatology. This encounter between African religions and biblical pneumatology reflects a genuine desire to make the doctrine of the Spirit relevant in an African context. This is supported by biblical texts, where such a 'dynamic pneumatology' is assumed.
Matthew Speaks To Ghanaian Healing Situations
Authors: Onyinah Opoku
Published: June 2007

This paper attempts to address the current approach to healing among Ghanaian Pentecostal/Charismatic churches. It traces the origins of the approach to the Ghanaian traditional practices, links it with the African Initiated churches and demonstrates that its current position was strengthened by the impact of the ministries of some evangelists in Ghana. Since there are some major problems with the current approach, lessons are drawn from Jesus's healing methods in the gospel of Matthew for the Ghanaian situations.
The Charismatic Structure of Acts
Authors: Thomas John
Published: June 2007

This study surveys major attempts to understand the structure of Acts and proposes a structure based upon insights gained from textual markers in the text and the significant role of the Holy Spirit. Building upon the work of a number of others, including Roger Stronstad in particular, this article proposes that the primary theme that holds the document together is the charismatic and/or the prophetic activity of the Holy Spirit. Accordingly, the proposal offered takes account of a full spectrum of defining emphases in Acts: the major individuals in Acts, the book’s emphasis upon missionary expansion, the literary markers in the text, and the strategic role of the Holy Spirit throughout the narrative.
Luminescence studies of the phagocyte response to endotoxin infusion into normal human subjects: multiple discriminant analysis of luminescence response and correlation with phagocyte morphologic changes and release of elastase
Authors: Taylor Fletcher , Haddad P.A. , Kinasewitz Gary , Chang Alvin , Peer Glenn , Allen Robert C.
Published: June 2007

A blood luminescence system (BLS) was employed to analyze blood phagocyte function in response to infusion of endotoxin (4 ng Escherichia coli lipopolysaccharide (LPS)/kg body weight) into 7 healthy human subjects. The subjects were closely monitored clinically, and extensive chemical, hematological and coagulation measurements were taken during the pretreatment, early (symptomatic, 1—8 h post-LPS), and late (asymptomatic, 12—48 h post-LPS) phases of acute inflammation. BLS assessment included measurement of basal and PMA-stimulated phagocyte oxidase and myeloperoxidase (MPO) activities, and also included measurement of circulating (COR) and PAF-primed maximum (MOR) opsonin receptor-dependent phagocytic activities. Basal oxidase activity peaked at T+1 h and showed an additional peak at T+24 h post-LPS. The COR activity also peaked at 1—2 h, but remained elevated through T+24 h post-LPS, while the basal MPO activity peaked only once at T+1 h. We concluded that while MPO evidence of phagocyte respiratory activation returned to baseline by T+ 4 h, COR evidence of receptor expression (receptor alert) remained elevated through T+24 h. During this early (0—8 h) period, elastase/α 1AT complex concentration peaked at T+3—4 h and again at T+8 h. Peak numbers of circulating polarized and vacuolated phagocytes also appeared at T+3 h and 7 h. We concluded that there was biochemical and morphological evidence of continuing phagocyte activity beyond T+4 h to T+8 h, and that this corresponded with the period during which the subjects were symptomatic. In addition, the appearance of a second peak of basal oxidase activity at T+24 h, multiple discriminant analyses of all the luminescence data, and the sustained elevation of lactate suggested that there was a later second stage (T+12 h to 48 h) of the human response to endotoxin, during which time the subjects were asymptomatic.
LPS and LAM activation of the U373 astrocytoma cell line: Differential requirement for CD14
Authors: Orr Sally L. , Tobias Peter
Published: June 2007

CD14 is a membrane protein (mCD14) found on monocytes and neutrophils that is required for the innate immune response to lipopolysaccharide (LPS) and lipoarabinomannan (LAM). CD14 can also be found in serum as soluble CD14 (sCD14) that when bound to bacterial products, enables many non-CD14 bearing cells to be activated. Lipopolysaccharide binding protein (LBP) is a plasma protein that disaggregates and catalytically transfers LPS to CD14. To examine the role of CD14 and LBP in LAM-dependent activation, we used the U373 astrocyte cell line to stably express membrane-bound CD14 (U373-CD14). In serum-free conditions, U373-CD14 cells could respond to LAM stimulation as measured by expression of intracellular adhesion molecule-1 (ICAM-1). Vector control cells (U373-RSV) could not respond to LAM or LPS; but, upon the addition of serum as a source of soluble CD14, control U373-RSV cells could respond to LPS, but not LAM. Therefore, LAM can activate U373 cells only through membrane CD14 and not soluble CD14. We also demonstrate that this membrane CD14-dependent LAM response is greatly facilitated by the addition of LBP.
The inhibitory action of butyrate on lipopolysaccharide-induced nitric oxide production in RAW 264.7 murine macrophage cells
Authors: Chakravortty Dipshika , Koide Naoki , Kato Yutaka , Sugiyama Tsuyoshi , Mu Mya Mya , Yoshida Tomoaki , Yokochi Takashi
Published: June 2007

The effect of butyrate, a natural bacterial product of colonic bacterial flora, on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 murine macrophage cells was studied. Butyrate significantly reduced NO production in LPS-stimulated RAW cells. The inhibition was abolished by the removal of butyrate. Butyrate also inhibited the expression of inducible type NO synthase (iNOS) in LPS-stimulated RAW cells. Furthermore, butyrate prevented the activation of nuclear factor (NF)-κB through the stabilization of IκB-α and IκB-β. Butyrate did not affect the phosphorylation of mitogen-activated protein (MAP) kinases by LPS. It was, therefore, suggested that butyrate down-regulated LPS-induced NO production in RAW cells through preventing the expression of iNOS, and that it was due to the inhibitory action of butyrate on the activation of NF-κB.
Endotoxin-mimetic effect of antibodies against Toll-like receptor 4
Authors: Nattermann Jacob , Xin Du  , Yongie Wei  , Shevchenko Dmitriy , Beutler Bruce
Published: June 2007

Monospecific, affinity-purified polyclonal antibodies reacting with the amino-terminal half of the mouse Toll-like receptor 4 (Tlr4) ectodomain failed to block LPS effects and, to the contrary, were capable of inducing TNF synthesis when applied to mouse macrophages and cross-linked with a secondary antibody. This effect was observed with macrophages derived from C3H/HeN and C57BL/10ScSn mice, but not with macrophages derived from C3H/HeJ or C57BL/10ScCr mice, indicating a specific, Tlr4-dependent effect. Neither primary nor secondary antibody caused any response if administered in the absence of the other reagent, nor was any response observed in cells from mice lacking Tlr4, or bearing the Lpsd mutation of Tlr4. These findings support several conclusions. Tlr4, the essential transducer of LPS responses, may act independently of LPS itself. LPS needs not be internalized, nor must it bind to a secondary target within the cell in order to exert its effect; rather, the receptor alone is required for initiation of a signal. The data are consistent with the hypothesis that a conformational change in Tlr4 is required for activation via this receptor, and reveal that the amino-terminal half of the Tlr4 ectodomain is a target sufficient for antibody-mediated activation.
Role of MD-2 in TLR2- and TLR4-mediated recognition of Gram-negative and Gram-positive bacteria and activation of chemokine genes
Authors: Dziarski Roman , Gupta Dipika
Published: June 2007

MD-2 is associated with TLR4 on the cell surface and enables TLR4 to respond to LPS. TLR2 without MD-2 does not respond to pure protein-free endotoxic LPS, ReLPS, and lipid A. MD-2 enables TLR2 to respond to non-activating LPS, ReLPS, and lipid A, and enhances TLR2-mediated responses to Gram-negative and Gram-positive bacteria, protein-containing LPS, peptidoglycan, and lipoteichoic acid. MD-2 enables TLR4 to respond to a wide variety of endotoxic LPS partial structures, Gram-negative bacteria, and Gram-positive lipoteichoic acid, but not to Gram-positive bacteria, peptidoglycan, and lipopeptide. MD-2 physically associates with both TLR4 and TLR2, but the association with TLR2 is weaker than with TLR4. Also, MD-2 and TLR2 and TLR4 enhance each other's expression. The highest induced genes in human monocytes stimulated with Gram-positive and Gram-negative bacterial cell wall components are chemokine genes, and IL-8 is the highest induced chemokine. Both Gram-positive and Gram-negative bacteria activate TLR2→ MyD88→IRAK→TRAF→NIK→IKK→NF-κB signal transduction pathway that induces transcription of the IL-8 gene. Therefore, TLR2 is a functional receptor for both Gram-positive and Gram-negative bacteria and it induces activation of IL-8.
Cytosolic protein ubiquitylation in normal and endotoxin stimulated human peripheral blood mononuclear cells
Authors: Majetschak Matthias , Suciu Dan Michael , Häsler Katrin , Obertacke Udo , Schade F. Ulrich , Jennissen Herbert P.
Published: June 2007

The ubiquitin-proteasome pathway is regarded as playing a crucial role in protein breakdown in inflammation and sepsis as well as in the regulation of inflammatory cell responses. In this pathway, ubiquitylation of target proteins is believed to act as a recognition signal for degradation by the 26S proteasome. As yet neither the ubiquitylation rate of cytosolic proteins, as a result of the total ubiquitin-protein ligase (tUbPL) activity, nor the specific ubiquitylation of calmodulin (ubiquitin-calmodulin ligase, uCaM-synthetase) has been determined in human mononuclear cells. Therefore, we studied cytosolic protein ubiquitylation in normal and in endotoxin (LPS)-stimulated human peripheral blood mononuclear cells (PBMNCs). PBMNCs from healthy volunteers were incubated with 0 or 100 ng/ml LPS for 18 h. Cytosolic extracts were obtained by hypotonic lysis and ultracentrifugation. TUbPL was measured as [125I]-[CT]-ubiquitin incorporation into the sum of cytosolic proteins. UCaM-synthetase activity was quantified with the fluphenazine (FP)-Sepharose affinity adsorption test. Endotoxin stimulation appears to inhibit tUbPL 3.7 ± 2.7-fold to 48 ± 43 fkat/mg (n = 6). UCaM-synthetase in cultures (n = 5) without endotoxin was determined to be 91 ± 32 fkat/mg +Ca2+ and 29 ± 23 fkat/mg —Ca2+. With endotoxin uCaM-synthetase was 138 ± 73 fkat/mg +Ca2+ and 14 ± 22 fkat/mg —Ca 2+. Ca2+-specificity (ratio ± Ca2+) of uCaM-synthetase increases from 3.1 without LPS to 10 after LPS stimulation, which was caused by a 2-fold decrease in minus Ca2+ activity and a 1.5-fold increase in plus Ca2+ activity. The data indicate specific regulatory effects of endotoxin on the cytosolic ubiquitylation systems in human PBMNCs.
Cellular activating properties and morphology of membrane-bound and purified meningococcal lipopolysaccharide
Authors: Bjerre Anna , Brusletto Berit , Rosenqvist Einar , Namork Ellen , Kierulf Peter , Øvstebø Reidun , Joø Gun-Britt , Brandtzæg Petter
Published: June 2007

Neisseria meningitidis, the cause of epidemic meningitis and acute lethal sepsis, synthesizes surplus lipopolysaccharides (LPSs) during growth, which are released as outer membrane vesicles (OMV) or `blebs'. Meningococcal disease severity is related to plasma LPS levels. We have compared the biological activities of native outer membrane vesicles (nOMV) to those of purified Nm-LPS (Nm-LPS) and LPS-depleted OMV (dOMV) prepared from N. meningitidis. The LPS content of nOMV was determined spectrophotometrically by quantifying KDO and by silver-stained SDS-PAGE gels. The morphology of the preparations was studied by transmission electron microscopy. The Limulus amoebocyte lysate (LAL) assay was used to quantify LPS in the plasma solutions. The preparations were diluted in endotoxin-free heparin plasma to equal amounts of LPS (w/w) in the range 50—5000 pg/ml. The biological reactivity was tested by: (i) a monocyte target-assay (monocyte purity ≥ 96%); and (ii) a whole blood model, measuring the secretion of TNF-α and IL-6 induction of procoagulant activity in monocytes (PCA). In both models, nOMV induced dose-dependent cell responses (TNF-α, IL-6, PCA) similar to purified Nm-LPS, whereas dOMV induced minimal responses. However, LAL activity was significantly higher for nOMV than for purified Nm-LPS and dOMV. The cellular responses of purified Nm-LPS and nOMV were reduced (>95%) by a specific anti-CD 14-antibody.
Examination of chlorpromazine and other amphipathic drugs on the activity of lipopolysaccharide antagonists, E5564 and E5531
Authors: Hua Yang  , Daun Jane M. , Rose Jeffrey R. , Christ William J. , Gusovsky Fabian , Chow Jesse C.
Published: June 2007

The synthetic antagonists of lipopolysaccharide (LPS), E5531 and E5564, are analogs of the lipid A portion of LPS that not only lack agonistic activity but also inhibit the biological effects of LPS both in vitro and in vivo. The effects of LPS and these synthetic antagonists have been localized to the recently described Toll-like receptor 4 (TLR4). A recent report indicated that the naturally occurring LPS antagonist Rhodobacter sphaeroides LPS loses its antagonist properties and gains pro-inflammatory qualities in the presence of chlorpromazine and other amphipathic drugs. To determine whether these reported actions occur with our chemically defined LPS antagonists, we examined the effects of chlorpromazine, fluphenazine, trifluoperazine, and lidocaine on the antagonism elicited by RsLPS and E5531 in U373 cells, which produce IL-6 in response to LPS. We also tested the effects of these amphipathic molecules on the LPS-neutralizing activity of RsLPS and E5564 on LPS-induced TNF-α release in human whole blood. The results indicate that neither chlorpromazine, fluphenazine, trifluoperazine nor lidocaine alter the activity of E5531 or E5564 in an in vitro cell system or human whole blood. Furthermore, chlorpromazine did not affect the antagonistic activity of RsLPS or E5564 on IL-6 generation by peripheral blood mononuclear cells. Thus, based on these data, our purified synthetic LPS-antagonists do not appear to lose their antagonistic properties and/or become agonists in the presence of amphipathic agents or drugs.
Mouse B1 cell line responds to lipopolysaccharide via membrane-bound CD14
Authors: Koide Naoki , Sugiyama Tsuyoshi , Kato Yutaka , Chakravortty Dipshika , Mya Mya Mu  , Yoshida Tomoaki , Hamano Teruaki , Yokochi Takashi
Published: June 2007

The role of membrane-bound CD14 in the response of mouse B1 cell lines to lipopolysaccharide (LPS) was studied. The surface profile of mouse TH2.52 B cells was positive for CD5, IgM, B220, CD11b and F4/80, suggesting that TH2.52 cells carried the typical phenotype of B1 cells. Furthermore, TH2.52 B1 cells were found to express membrane-bound CD14, which plays a critical role in LPS recognition. TH2.52 B1 cells responded to a very low concentration of LPS and exhibited: (i) augmentation of DNA synthesis; (ii) activation of nuclear factor (NF)-κB; and (iii) phosphorylation of extracellular signal regulated kinase 1/2 (Erk1/2). They were markedly inhibited by anti-CD14 antibody. Therefore, the expression of membrane-bound CD14 was suggested to provide high sensitivity to LPS for TH2.52 B1 cells.
Review: Immunodepression in sepsis and SIRS assessed by ex vivo cytokine production is not a generalized phenomenon: a review
Authors: Cavaillon Jean-Marc , Adib-Conquy Minou , Cloëz-Tayarani Isabelle , Fitting Catherine
Published: June 2007

Sepsis and non-infectious systemic inflammatory response syndrome (SIRS) are paradoxically associated with an exacerbated production of cytokines, as assessed by their presence in biological fluids, and a diminished ability of circulating leukocytes to produce cytokine upon in vitro activation. In this review, we depict that the observed cellular hyporeactivity is not a global phenomenon and that some signalling pathways are unaltered and allow the cells to respond normally to certain stimuli. Furthermore, we illustrate that during sepsis and SIRS, cells derived from tissues are either fully responsive to ex vivo stimuli or even primed, in contrast to cells derived from hematopoietic compartments (blood, spleen, etc.) which are hyporeactive. In addition to cytokine production, nuclear factor-κB (NF-κB) status within leukocytes can be used as a useful marker of hypo- or hyper-reactivity. We illustrate that the immune-depression reported in sepsis and SIRS patients, often revealed by a diminished capacity of leukocytes to respond to lipopolysaccharide, is not a generalized phenomenon and that SIRS is associated with a compartmentalized responsiveness which involves either anergic or primed cells.
IC14, a CD14 specific monoclonal antibody, is a potential treatment for patients with severe sepsis
Authors: Axtelle Tim , Pribble John
Published: June 2007

CD14 is a pattern recognition receptor for the bacterial cell wall components from Gram-positive and Gram-negative bacteria as well as mycobacteria. Binding of lipopolysaccharide (LPS) or other cell wall constituents to CD14 initiates signal transduction through the Toll-like receptors resulting in the release of pro-inflammatory cytokines and the initiation of the systemic inflammatory response. In rabbits and non-human primates, CD14 specific antibodies were shown to attenuate responses to LPS or Escherichia coli challenge including pro-inflammatory cytokine release, acute lung injury, hypotension and changes in lung, liver, spleen and adrenal gland morphology. In healthy human subjects, single doses of a chimeric CD14 antibody (IC14) have been shown to be well tolerated and result in a dose-related degree of saturation of CD14 receptors on monocytes and granulocytes. Pretreatment of healthy subjects with IC14 2 h prior to LPS resulted in an attenuation of the LPS-induced fever, clinical symptoms, and leukocyte activation and degranulation. IC14 inhibited the release of TNF-α, IL-6, and IL-10 and delayed the release of sTNFRI and IL-1ra. Further studies are in progress to characterize the safety and clinical pharmacology of IC14 in patients with severe sepsis.
Lipopolysaccharide stimulates HIV-1 entry and degradation in human macrophages
Authors: Pushkarsky Tatiana , Dubrovsky Larisa , Bukrinsky Michael
Published: June 2007

The bacterial endotoxin LPS is a potent stimulator of monocyte and macrophage activation and has been shown to protect differentiated macrophages from de novo infection by HIV-1. However, the mechanisms of this inhibitory activity of LPS are not fully understood. We investigated the effect of LPS on the early post-binding steps of HIV-1 replication in primary macrophages. Paradoxically, when applied together with the virus, LPS stimulated entry of HIV-1 into macrophages, as judged by the amount of internalized HIV-1 RNA and p24. This stimulatory activity did not depend on receptors used for entry and did not require new protein synthesis. However, internalized viral RNA and p24 were rapidly degraded in LPS-stimulated macrophages. Surprisingly, while degradation of HIV-1 p24 in LPS-treated cells was inhibited by bafilomycin A1, HIV-1 RNA was not protected by this agent, suggesting that viral RNA is degraded by a pH-independent mechanism. These results indicate that LPS stimulates both virus uptake and virus degradation in macrophages.
Vagotomy attenuates the effect of lipopolysaccharide on body temperature of rats in a dose-dependent manner
Authors: Azab Abed N. , Kaplanski Jacob
Published: June 2007

There is a growing body of evidence suggesting that vagal afferents play a major role in peripheral-neural communication. This study was undertaken to determine whether a dose-dependent effect of lipopolysaccharide (LPS) on vagotomy-induced febrile unresponsiveness exists, and to examine the effect of vagotomy on LPS-induced increase in hypothalamic prostaglandin E2 (HT PGE2) production. Vagotomized and sham-operated rats were subjected to two experimental protocols. In the first, vagotomized and sham-operated rats were injected intraperitonealy with different doses of LPS (200, 500 and 1000 µg/kg) in order to examine the dose-dependent effect of LPS on the biphasic febrile response of the rats. In the second protocol, vagotomized and sham-operated rats were injected intraperitonealy with LPS (500 µg/kg). Two hours post injection, body temperature was measured, the rats were decapitated and blood was collected. Simultaneously, the rats' hypothalami were excised and incubated for 1 h in a Krebs-Henseleit buffer. Next, HT PGE2 was determined by radioimmunoassay. Vagotomy-induced gastric enlargement was then measured to examine the correlation between the magnitude of the enlargement and that of the vagotomy-related febrile unresponsiveness. It was found that vagotomized-induced febrile unresponsiveness is a dose-dependent effect. Subdiaphragmatic resection of the vagus prevented the biphasic febrile response caused by the lowest dose (200 µg/kg) of LPS, whereas the highest dose of LPS (1000 µg/kg) caused a similar biphasic febrile response in both vagotomized and sham-operated rats. Indeed, vagotomy attenuates LPS-induced increase in HT PGE2, and blocks the hypothermic phase of the febrile response. On the other hand, no correlation between gastric enlargement and febrile unresponsiveness was found. The results of the present study may cast further light on the crucial role of the vagus nerve as a peripheral-neural pathway in the mediation of the febrile response.
Genetic determinants of lipopolysaccharide and D-galactosamine-mediated hepatocellular apoptosis and lethality
Authors: Oberholzer Andreas , Oberholzer Caroline , Bahjat F.R. , Edwards Carl K. , Moldawer Lyle L.
Published: June 2007

Lipopolysaccharide and D-galactosamine induced lethality and apoptotic liver injury is dependent upon endogenously produced TNF-α. Unlike the response to high dose lipopolysaccharide alone, death in this model is a direct result of hepatocyte apoptosis. In a series of recent studies, we have demonstrated that mortality and hepatic injury following lipopolysaccharide administration in D-galactosamine-sensitized mice is dependent upon secreted 17 kDa TNF-α acting primarily through the p55 TNF receptor. Transgenic mice expressing null forms of TNF-α, the p55 receptor, or expressing only a cell-associated form of TNF-α exhibited no mortality and only modest liver injury when challenged with 8 mg of D-galactosamine and 100 ng of lipopolysaccharide. Although Fas ligand expression is increased in the liver, it appeared to play no significant role in outcome, since mice expressing a mutant form of Fas ligand are still sensitive to LPS- and D-galactosamine-induced lethality. Finally, we have seen significant variation in LPS- and D-galactosamine-mediated lethality among different strains of mice. The non-obese diabetic or NOD mouse is highly resistant to LPSand D-galactosamine-induced lethality, and this appears to be secondary to a post-receptor defect in p55 TNF receptor signaling. The studies confirm an essential role for TNF-α and p55 TNF receptor signaling in the hepatocyte apoptosis and lethality associated with lipopolysaccharide and D-galactosamine administration.
Blockade of CD14 aggravates experimental shigellosis
Authors: Wenneras Christine , Ave Patrick , Huerre Michel , Arondel Josette , Ulevitch Richard , Mathison John , Sansonetti Philippe
Published: June 2007

Shigella infections lead to severe inflammation associated with destruction of colonic mucosa. We assessed the effect of in vivo blockade of CD14 on the outcome of experimental Shigella infection in rabbits. A total of 17 rabbits were divided into two groups: 8 received a single i.v. dose of anti-rabbit CD14 monoclonal antibody prior to infection with an invasive Shigella flexneri strain; the remainder served as controls. The anti-CD14-treated rabbits exhibited more severe tissue destruction and a 50-fold increase in bacterial invasion of the intestinal mucosa when compared to controls. Similar numbers of polymorphonuclear leukocytes were recruited to the intestinal mucosa in both groups despite the massive bacterial invasion seen in the CD14-blocked group. No statistically significant differences were seen in levels of IL-1β nor in the ratio of IL-1RA/IL-1β for either group. In contrast, higher quantities of TNF-α were observed in the CD14-blocked group. To conclude, anti-CD14 treatment had a detrimental effect on the capacity of Shigella-infected animals to clear the infection.
A novel strategy for the synthesis of neoglycoconjugates from deacylated deep rough lipopolysaccharides
Authors: Müller-Loennies Sven , Grimmecke Dieter , Brade Lore , Lindner Buko , Kosma Paul , Brade Helmut
Published: June 2007

We report a novel strategy for the preparation of neoglycoconjugates of oligosaccharides which are obtained after complete deacylation of bacterial deep rough lipopolysaccharides (LPS) isolated from recombinant Escherichia coli bacteria synthesizing a Kdo di-[α-Kdo-(2→4)-α-Kdo-(2→] and a Kdo trisaccharide [α-Kdo-(2→8)-α-Kdo-(2→4)-α-Kdo-(2→] of Re-type and chlamydial LPS, respectively. Unlike acylated LPS, such oligosaccharides can be obtained in pure form and thus lead to well-defined neoglycoconjugates. Cleavage of the 1-phosphate of the lipid A moiety by alkaline phosphatase treatment leads to a free reducing glucosamine which can be further reacted with allylamine. After reductive amination, spacer elongation of the allyl group with cysteamine and activation with thiophosgene, the ligands were reacted with BSA. We have compared the immunological reactivity of such defined neoglycoconjugates obtained from natural sources with those obtained by chemical synthesis and report that such neoglycoconjugates are immunogenic and well suited as antigens for the study of epitope specificities of monoclonal antibodies. In addition, we have compared these conjugates with those in which ligands were coupled by glutardialdehyde to BSA. Our approach proved to be superior since the latter led upon immunization of mice to a relatively high percentage of antibodies that reacted with glutardialdehyde derivatized BSA without the carbohydrate ligand. This was not the case for cysteamine-spacered ligands coupled via their isothiocyanate-derivatives.