bluuebunny/crossref_metadata_2025 huggingface dataset:
Administrative Culture and Policy Making in The Department of the Environment
Authors: Wilks Stephen
Published: January 2008

The article explores the potenztialfor developing a concept of administrative culture and identifying its contribution to policy-making within central government. It takes the case of the DOE and analyses the influence offunction, clientele, resources and history on culture. It goes on to argue that this culture is functional anld that the policies of recent Conservative Governments, especially in the area of urban policy, which have cut across such norms, hlave encountered administrative opposition and a predictable implementation deficit. Examples are drawn from judicial judgements on housing and local governmenit finance. The article concludes thlat the administrative culture of the DOE impedes urbani policy but retainis some laudable features.
Bigger Business and the State
Authors: van Tulder Rob
Published: January 2008

This article considers the role of Government in the support of high technology innovation. In particular it draws attention to the difference between Governmental relations with large firms and small firms. It argues that while rhetoric might give prominence to the support of small companies, there are in-built pressures to support ‘national champions.’
Trading work: discourses of internal exchange in the economic culture of local government
Authors: Mackintosh Maureen
Published: January 2008

Local authorities increasingly use ‘trading’ language to describe and debate inter and intra-departmental working relations. This article explores the meanings given to this contested language in context, drawing on case studies of internal trading in two authorities. The objective of this interpretative exercise in economic discourse is to explore the extent to which a dominant discourse of internal ‘trading’ exists within the two authorities, and its association with identifiable patterns of economic behaviour. The article identifies four discourses of internal ‘exchange’, of which two, labelled the ‘trading’ and the ‘public business' discourses, are widely shared and attempt to ally social and public concern with devolved financial management to business-like behaviour. Each of these two discourses was identified with distinct policies and preferred behaviour in internal working relations. Neither was ‘winning’: we are a long way from the solidifying of a single new internal economic culture in local government through the language of trade.
The Privatisation of the Property Services Agency: Risk and Vulnerability in Contract-Related Fraud and Corruption
Authors: Doig Alan
Published: January 2008

The Property Services Agency was a ‘hived-off’ government organisation responsible for the design, building, renting and maintenance of government property and associated facilities. Its expenditure, and delivery of its functions, were criticised for fraud, waste and inefficiency while it was involved in a number of cases of contract-related fraud and corruption.

As the Property Services Agency began to deal with the latter concerns, the government was making it a prime candidate for a purchaser-provider split in relation to funding, the divestment of a number of functions to other agencies and to its client departments, and the introduction of competition in the provision of its services. Finally the Property Services Agency became a candidate for privatisation as its contracting functions were fully transferred to government departments. The responsibilities for combatting corruption and fraud, however, have also been dispersed, leaving HM Treasury to report its concerns over the threat such an area of risk area poses for government departments who have wanted the responsibility for the Agency's functions but, without the awareness, expertise and economies of scale to respond effectively, may be vulnerable to contract-related fraud and corruption. The functions of the Agency, its privatisation, and the implications for dealing with contract-related fraud and corruption provide a case-study in the relationship between different forms of organisation and ownership of the operational and purchasing functions of government, and the ability of governments to control corruption and fraud in public contracting.
Markets, Quasi-Markets and Middle Managers in Local Government
Authors: Keen Linda
Published: January 2008

Operating as purchasers or providers, middle managers play a key role in the implementation of the new market systems recently introduced within local authorities. However, relatively little empirical investigation has been undertaken into the first-hand experiences of the middle managers who are on the ‘receiving end’ of these market systems. After a brief review of the literature, this paper uses Williamson's (1975; 1985) model of behavioural attributes as an analytical framework for analysing the impact of the new Purchaser and Provider model on the performance of middle line managers working within a local authority well-known for its commitment to market service delivery systems.

These managers' responses about the positive and negative aspects of their experiences of market systems varied significantly according to their roles as either Purchaser or Provider managers, and to their location within particular service departments. The paper draws attention to the important differences in the nature of the transactions within each of these departments which appeared to account, at least in part, for the managers' different responses, and argues against the wholesale application of market forms of service delivery across highly differentiated organisations such as local authorities.
Incrementalism and Rationalism in Local Government Reorganisation 1957-1996
Authors: Leach Robert
Published: January 2008

This article seeks to apply rationalist and incrementalist models of the policy process to local government reorganisation from 1957 to 1996. The ambiguities and tensions in the models between description and prescription and between process and outcome are briefly explored, and the complexity of the models acknowledged, as a prelude to their application to the wide variety of approaches to the structural reorganisation of local government. While the quasi-incremental processes of the 1958 and 1992 Local Government Commissions are widely perceived to have failed, most of the recommendations of the more ostensibly rational Herbert, Wheatley and Redcliffe-Maud Royal Commissions have not endured, suggesting some inherent problems with both models. The frequency and extent of changes in local government structure scarcely confirm incrementalist expectations over outcomes, yet the failure of the more radical innovations to win long-term acceptance and legitimacy may be held to give some retrospective justification for gradualist caution. However, the cumulative failure of the reform process may ultimately reflect a misdiagnosis of the underlying problems of local government - essentially a rationalist critique. If a defective structure was not the main problem, reorganisation was not the solution.
Strategic change in Government Agencies influenced by Apollo and Zeus-two ‘Gods of Management’
Authors: McHugh Marie
Published: January 2008

This paper argues that many Next Steps agencies have been forced to embark upon far reaching programmes of strategic change in an attempt to satisfy externally imposed demands for enhanced levels of efficiency and effectiveness. However, it would seem that the management of the entire change process has been problematic, and has been characterised by disintegration in the form of a series of disjointed organisational initiatives. Focusing upon the experience of one Next Steps agency, it is argued that a root cause of the problem may lie in a rigid adherence to an outmoded value system symbolised by allegiance to ‘Apollo’ the Greek god of order. Thus, to effect successful strategic change within government agencies it is suggested that they must, in the first instance, seek to foster allegiance to a new and more dynamic figure in the form of the Greek god ‘Zeus’.
Contracting for Services-a Double Jeopardy? An Analysis of Contract Compliance in the Context of European and UK Social and Public Procurement Policy
Authors: Fee Ruth , Maxwell Patricia , Erridge Andrew
Published: January 2008

This article discusses the scope for implementing a contract compliance policy in the UK as part of the new Best Value regime for local authority contracting. The article examines the interrelationship between compulsory competitive tendering policy in the UK and EU social and public procurement policies, drawing upon secondary research and case-law on the social and employment effects of contracting-out. The article identifies the problem of a ‘double jeopardy’ for local authorities adopting contract compliance policies: even if national procurement policies are adapted to accommodate these policies, any attempt to impose contract compliance through large service contracts will be contrary to EU law under the services directive.
Whitehall in the 1950s and 1960s
Authors: Fry Geoffrey K.
Published: January 2008

This paper was originally presented at the ESRC Whitehall Programme/Public Record Office Conference on Whitehall in the 1950s and 1960s, Policy Making in an Expanding Government, which was held at the Public Record Office, Kew on April 16-1 7th 1997, and it is now published in a form that has been subject to the refereeing process. Its argument is that in the first part of the 1950s, as had been the case really since 1945, the dominant attitude towards British central government administrative arrangements - Whitehall, in shorthand terms - was Panglossian, meaning that the prevailing wisdom was that ‘we lived in the best of all possible worlds.’ After the trauma of Suez in 1956, this mood changed to what at times seemed to amount to an obsession with recasting the organization and staffing of Whitehall departments, as if ‘we lived in the worst of all possible worlds.’ In the spheres of defence and external affairs, the changes made tended to have more logic to them than was evident over in other areas of public policy, where the main objective seemed to be to shore up the Keynesian Welfare State, unless, that is, it was to give an impression of activity on the part of politicians.
In what sense a National Health Service?
Authors: Powell Martin
Published: January 2008

Under the recent Conservative and current Labour Governments it appears that the NHS is moving in two different directions at once. Some commentators claimed that the national character of the health service was undermined while others argued that the NHS was effectively nationalised. One reason for these apparently contradictory positions is that there are no clear criteria of what a National Health service is. This article presents three main characteristics of a national health system, with illustrations from the NHS. Recent events seem to confirm that the NHS has never been clearly a national or a local service, and existing trends seem to lead to the worst of all worlds: the disadvantages of central control, and local differentiation without any genuine local autonomy. As the NHS is arguably the most national service in the British welfare state, it is possible that the heyday of the national welfare state may be over.
A collocational approach to semantic change: the case of worship and honour in Malory and Spenser
Authors: McBride Christopher
Published: January 2008

The word worship, which in Late Middle English named an obligation-related social value, had by early modem times been largely replaced with honour. This article uses collocational data from two literary texts - Malory's Works and Spenser's The Faerie Queene - to propose an explanation for this change. Patterns of lexical connection for worship and honour support their respective assignment to two different social paradigms, status and contract. The different semantic categories which are present in the collocational data suggest that the change from worship to honour may be part of a larger rearrangement of the vocabulary of obligation in the period, caused by the contemporary shift of dominant social paradigm from status to contract. A sociohistorical analogue to the linguistic case is also presented.
Basil Bunting's Briggflatts : a case study in intonational prosody
Authors: Pople Ian
Published: January 2008

In this article I wish to contribute to the analysis of prosody in poetry by looking at free or 'unmetred' verse. In particular I focus on the way in which the tools for analysing intonation may be used for analysing the performance of poetry in order to examine the way lineation is formed in unmetred verse. I look at the way tone-unit boundaries are often co-extensive with line endings in unmetred verse. The article follows work in this kind of analysis by Crystal (1975) and concentrates on the poem Briggflatts by Basil Bunting. Syntactic features such as ellipsis and pre- and post-modification effect the placement of tone-unit boundaries. Line endings may also be effected by the poet's use of marked emphases, and by the influence of other prosodies which a sophisticated poet may bring to bear on his material.
Monoclonal antibodies against the heptose region of enterobacterial lipopolysaccharides
Authors: Swierzko A. , Brade L. , Brabetz W. , Zych K. , Paulsen H. , Brade H.
Published: January 2008

Murine monoclonal antibodies (MAbs) against the rough mutant lipopolysaccharides (LPS) of Salmonella minnesota strain R4 and R7 (chemotypes Rd2P - and Rd1P-, respectively) were obtained after immunization of BALB/c and NZB mice, respectively, with heat-killed bacteria. A total of 5 MAbs was obtained which were serologically characterized by hemagglutination, EIA and Western blot using LPS, de-O-acylated LPS and dephosphorylated LPS. In addition, the completely deacylated carbohydrate backbone, the deacylated, dephosphorylated and reduced carbohydrate backbone, and synthetic partial structures were covalently linked to bovine serum albumin resulting in artificial glycoconjugate antigens. Both groups of MAbs (anti-R4 and anti-R7) were highly specific for the Rd2P- and Rd1P- chemotypes, respectively, since they did not react with LPS of the Ra, Rb, Rc or Re chemotype. The R4 antibodies required for binding the core region [composed of 3-deoxy-α-D- manno-octulosonic acid (Kdo) and L-glycero-α-D- manno-heptopyranose (Hep)] and the phosphorylated glucosamine backbone of the lipid A moiety. The R7 antibodies bound to some extent to the core oligosaccharide Hep(1→3)Hep(1→5)Kdo(2→6), however, better binding was observed when the glucosamine backbone of lipid A was also present. In this case however, phosphoryl groups were not required. These antibodies can detect enterobacteria with an LPS of the Rd-chemotype, and may be useful to characterize also other naturally occurring rough mutants or genetically manipulated strains used to clone genes of the rfa locus.
Minireview: Therapeutic intervention in sepsis with antibody to endotoxin: is there a future?
Authors: Cross A.S. , Opal S.
Published: January 2008

Within the last 24 months a number of studies that tested the efficacy of immunologic reagents in the treatment of sepsis were concluded. Among these reports were 4 studies1-4 completed on 2 anti-endotoxin monoclonal antibodies (MAb, HA-1A and E-5). These clinical trials did not generate data sufficient to support product licensure. Given the attention and expectations surrounding the anti-endotoxin MAbs, the disappointing results raised the question whether the use of anti-endotoxin antibodies in the treatment of sepsis was still a viable concept.5 The question was rendered even more relevant given the decade-old controversy surrounding the efficacy of polyclonal antibodies to endotoxin, particularly antibody to the J5 (Rc chemotype) mutant of Escherichia coli 0111:B4, a conceptual progenitor of the HA-1A and E-5 MAbs.6 In this review we shall examine whether anti-endotoxin antibodies may yet offer any therapeutic potential in the treatment of sepsis. It will be our contention that antibodies to core glycolipid will be useful adjuncts to therapy, particularly if used as part of combination immunotherapy.
Elimination of trace endotoxin protein from rough chemotype LPS
Authors: Manthey C.L. , Vogel S.N.
Published: January 2008

The phenol-chloroform-petroleum ether (PCP) extraction is the method of choice for isolating LPS chemotypes from rough mutant bacteria. However, we have observed that a high percentage of PCP-purified LPS preparations contain trace levels of protein contaminants, and these protein contaminants may exhibit endotoxin-like activity. To obtain protein-free rough LPS, a modified phenol-water extraction procedure was developed for use as a final step to follow PCP extraction. Using this procedure, Salmonella minnesota Ra, Rc, Rd and Re LPS were recovered in yields of 82-100% from the original PCP-extracted preparations. Yields were determined by recovery of KDO and supported by analysis of silver stained SDS-PAGE gels. Although the original PCP-purified chemotypes were contaminated by major 41, 38.5, 29.5, 23, 17 and 14 kDa proteins (detected by Western blotting and gold staining), the repurified LPS contained no detectable protein. The repurified LPS retained all of its original bioactivity as defined by the ability to stimulate normal C3H/OuJ macrophages to secrete TNF. The importance of utilizing repurified LPS in biologic studies was illustrated by the loss of > 99% of activity when repurified LPS was used to stimulate TNF secretion by LPS 'hyporesponsive' C3H/HeJ macrophages.
Endotoxemia-related attenuation of interleukin-1 secretion in thermally injured patients
Authors: Cembrzynska-Nowak M. , Lalani S. , Peters W.J. , Teodorczyk-Injeyan J.A.
Published: January 2008

The present study examines the effect of post-burn endotoxemia on the capacity for the in vitro induction of interleukin-1 (IL-1)β synthesis by either inflammatory or non-inflammatory stimuli. Adherent cell (AC) cultures from patients with severe burns (n = 14, 25-65% total body surface area) were stimulated with LPS, phorbol ester (TPA) or IL-2 and studied simultaneously with intact or LPS-pretreated preparations from normal donors (n = 9). Levels of spontaneously released IL-1β in unstimulated AC cultures from patients and in LPS-pretreated preparations from normal individuals were highly elevated. However, LPS- and IL-2-related IL-1β production in such cultures was significantly decreased (P < 0.01-0.05). In contrast, in the majority of patients' cultures and LPS-pretreated normal AC preparations, TPA-induced IL-1β production remained relatively unaffected. In all patients studied, reduced capacity for the LPS-induced IL-1 production was observed concurrently with a significant (P < 0.05) elevation in plasma endotoxin content. Diminished secretion of IL-1β coincided also with a profound reduction of IL-2-dependent IFNγ production in patient cultures and in LPS-pretreated normal cultures. Thus, in the burn patient, endotoxemia-related specific desensitization may develop and, combined with other intrinsic mechanisms, restrict the pathophysiological sequelae of continuous bacterial stimulation.
Mitogenicity of bacterial lipopolysaccharide on the T lymphocyte population bearing the γδT cell receptor
Authors: Nitta T. , Imai H. , Ogasawara Y. , Nakano M.
Published: January 2008

In vitro [3H]-thymidine (TdR) uptake in T lymphocyte populations, taken and purified from proteose peptone-induced peritoneal exudate cells (PEC), spleens and thymuses of C3H/HeN mice, were investigated by stimulation with bacterial lipopolysaccharide (LPS). The T cells taken from PEC (PEC-T) were purified by passing them through a nylon fiber column (Nfc) and a G-10 column. They were then treated with anti-lak, anti-Mac-1 and anti-LR-1 antibodies plus complement (C). The PEC-T cells showed an increased [ 3H]-TdR uptake in response to LPS, while the splenic T cells and thymic T cells that had been purified and treated by a similar procedure with minor modifications, poorly responded to LPS and did not respond to LPS, respectively. The response of PEC-T to LPS was abolished when the PEC-T were pretreated with anti-Thy-1 or anti-γδ antibody plus C, but not αβ, T cell receptor (TCR) antibody plus C. The thymic T cells did not show any increase of [ H]-TdR uptake in response to LPS or anti-αβ or γδTCR antibody. However, obvious uptake did occur when the cells were stimulated with LPS and anti-γδTCR antibody, but not with anti-αβTCR antibody. IL-1 did not substitute for LPS in the response. These results suggest that LPS has a mitogenic ability to respond to a T cell population bearing γδTCR in PEC, and to a γδTCR-stimulated cell population in thymic cells. The difference in response to LPS between PEC-γδT and thymic γδT cells was discussed.
Population dynamics of inducible nitric oxide synthase production by LPSand LPS/IFNγ-stimulated mouse macrophages
Authors: Pace J.L. , Lowenstein C.J. , Phillips T.A. , Chen L.C. , Morrison D.C. , Hunt J.S. , Russell S.W.
Published: January 2008

The reactive nitrogen intermediate, nitric oxide (NO) is important in host defense against both NO-sensitive microorganisms and tumor cells. Macrophages are one of the chief inflammatory sources, especially when stimulated with the combination of LPS and interferonγ (IFNγ). It is not known, however, whether IFNγ-mediated augmentation of LPS-induced production of NO is the result of greater production by all cells or to the recruitment of more producer macrophages within a given population. This question was addressed, first, by stimulating mouse macrophages (either bone marrow culture-derived, inflammatory peritoneal or those of the cell line, RAW 264.7) with up to 10 U/ml IFNγ for as long as 24 h. Under these conditions, there was little or no production of NO and rare or no cells were immunocytochemically positive for the inducible form of nitric oxide synthase (iNOS), which catalyzes the production of NO. Populations similarly exposed to 1 ng/ml LPS were low producers of NO and contained somewhat more, but still only a few (< 15%), iNOS-positive cells. In contrast, as the concentration of IFNγ was increased (≥ 1 U/ml) in the presence of a constant amount of LPS (1 ng/ml), the principal effect was to increase both the production of NO and the number of iNOS-positive macrophages. The amount of iNOS expressed by some cells also appeared to be increased. Two important conclusions can be drawn from these findings: (1) there is heterogeneity in mouse macrophage populations with respect to the production of iNOS; and (2) increasing concentrations of IFNγ appear to augment LPS-induced secretion of NO by recruiting increasingly greater numbers of macrophages into the production of iNOS. Such results potentially provide important clues as to how IFNγ may be acting at the subcellular level to enhance iNOS synthesis.
Human hemoglobin increases the biological activity of bacterial lipopolysaccharides in activation of Limulus amebocyte lysate and stimulation of tissue factor production by endothelial cells in vitro
Authors: Kaca W. , Roth R.I. , Ziolkowski A. , Levin J.
Published: January 2008

Previous studies have demonstrated that hemoglobin (Hb) and bacterial endotoxin (lipopolysaccharide, LPS) form stable complexes and result in disaggregation of macromolecular LPS. To examine the effect of complex formation on LPS biological activity, we investigated the ability of Hb to alter LPS-induced activation of the coagulation cascade of Limulus amebocyte lysate (LAL) and expression of tissue factor from human endothelial cells. Both native HbAo and derivatized (covalently cross-linked) hemoglobin resulted in prominent enhancement of LAL activation and endothelial cell tissue factor production by Proteus mirabilis LPS. No substantial differences were observed between the enhancement effect of Hb on P. mirabilis smooth and rough LPS, indicating a dominant role for the lipid A component of LPS. Rough (Re) Salmonella min nesota 595 LPS also demonstrated both enhanced activation of LAL and stimulation of endothelial cell tissue factor in the presence of Hb. In contrast, neither lipid A nor singly dephosphorylated or partially deacylated Re LPS manifested significant enhancement of LAL activation by Hb, and partially deacylated Re LPS showed no enhancement of endothelial cell tissue factor by Hb. These results suggest that the Kdo moieties, as well as the phosphate residues and fatty acyl moieties of lipid A, may be involved in the interaction of Hb with LPS. Comparison of Hb with other endotoxin binding proteins for ability to cause enhancement of LPS biological activity demonstrated more prominent enhancement with lipopolysaccharide binding protein (LBP) than that observed with Hb, lesser enhancement with albumin, and no enhancement effect with IgG or transferrin.
Carnitine and septic shock: a review
Authors: Famularo G. , De Simone C. , Arrigoni Martelli E. , Jirillo E.
Published: January 2008

Most studies have reported reduced carnitine levels in the tissues of patients with sepsis, probably due to increased urinary excretion. Because of the increased utilization of fatty acids and ketone bodies as sources of energy in sepsis, the carnitine deficiency can further impair the fuel metabolism and contribute to the unregulated lipid metabolism in these patients.

Recently, experimental and clinical studies have shown that carnitine and its congeners are able to: (a) downmodulate the spontaneous and endotoxin (LPS)-triggered overproduction of tumor necrosis factor (TNF)-alpha; (b) ameliorate the lipid metabolism; and (c) reduce the severity of illness, accelerate recovery, and, in some cases, improve survival in experimental septic shock.

Many questions concerning the ultimate molecular mechanism of action of these compounds in endotoxaemia are still unanswered. Yet, these compounds may be helpful in patients with sepsis, when associated with conventional therapy, in that they can effectively reduce TNF-alpha levels and ameliorate the host's metabolic processes.
Lessons learned from clinical trials of sepsis
Authors: Opal S.M.
Published: January 2008

The development of novel therapeutic agents with demonstrable clinical efficacy in septic patients has proven to be an elusive goal. Despite a biologically plausible therapeutic rationale, antisepsis drugs have not performed consistently or convincingly in recent clinical trials. A great deal of information has been learned recently about the basic mechanisms and molecular pathophysiology of sepsis. Nonetheless, the optimal patient population, the clinical and laboratory indicators to predict outcome and susceptibility to treatment, and the most appropriate clinical endpoints have yet to be clearly defined. Refinements in clinical trial design may be as important as basic discoveries in molecular biology in the future development of new therapeutic agents in septic shock.
Can second-generation versions of existing therapeutics be successful in clinical trials of sepsis?
Authors: Fisher C.J.
Published: January 2008

Sepsis syndrome and septic shock remain as significant causes of morbidity and mortality. To date, clinical trials of immunotherapeutic agents have failed to establish conclusively a role for these agents despite considerable animal data to suggest a positive benefit. This article discusses the hypotheses upon which these therapies are based, the critical issues associated with clinical trials and the molecules which have been clinically tested. Recommendations for future clinical trials in sepsis are made.
Physicochemical and biological characterizations of complex formation between lipopolysaccharide and lysozyme
Authors: Takada K. , Ohno N. , Yadomae T.
Published: January 2008

Endotoxin (lipopolysaccharide, LPS) from Gram-negative bacteria induces a variety of biological effects in mammals. We have earlier demonstrated that lysozyme (LZM) will bind to both smooth and rough forms of LPS and lipid A with high affinity, and inhibits several of the immunomodulatory activities of LPS. The production of LZM/LPS complexes was suggested to be significantly influenced by both reaction temperature and ionic strength. In this manuscript, we describe experiments designed to examine the formation of LZM/LPS complexes prepared under various conditions as assessed by fluorescence measurements of dansylated LZM and also by separation of complexes via nitration methodologies. The results of these studies provide additional experimental evidence that complex formation requires both ionic and hydrophobic interactions. The most stable complexes were formed at conditions of low ionic strength and higher temperature. Under these conditions, hydrophobic interactions were predominant in complex formation. At lower temperatures, the extent of complex formation was less, and ionic interactions were predominant. TNF production by RAW264.7 cells in vitro was decreased by LZM/LPS complexes prepared in both high and low ionic strength, but was most significantly decreased at conditions of low ionic strength and higher temperatures. These facts suggest that both ionic and hydrophobic interactions of LZM with LPS are important for neutralizing the biological activity of LPS.
Lipopolysaccharide is required for the lethal effects of enterotoxin B after D-galactosamine sensitization
Authors: Chapes S.K. , Beharka A.A.
Published: January 2008

We tested the D-galactosamine sensitization model with staphylococcal enterotoxin B (SEB). LPS was required for the lethal effects of SEB in D-galactosamine sensitized mice. Only two (2/62) among the C3HeB/FeJ (H-2k), Balb/c (H-2d) and C57BL/6J (H-2b) mice died in response to SEB in the absence of LPS whereas injection of SEB and minimally lethal concentrations of LPS became highly toxic. Similar to LPS, the lethal effect of SEB was dependent on the mouse strain used. Mouse strains more sensitive to the effects of LPS (Balb/c and C57BL/6J) were also more sensitive to the effects of SEB in comparison to C3H mice when equivalent doses of LPS and SEB were used. Among Balb/c and C3HeB/FeJ but not C57BL/6J mice, SEB (20 μg) potentiated the lethal effects of LPS at low doses (0.1 μg LPS), but had an apparent protective effect at high doses (1 μg LPS). Lastly, there was an inverse correlation between pathogenesis and serum IL-2 concentrations and splenic T cell activation in C3H mice. However, macrophage mobilization did correlate with lethality. Therefore, some questions remain about the mechanisms involved in the D-galactosamine/SEB pathogenesis model. We conclude that when sensitizing mice with D-galactosamine and assessing the lethal effects of SEB, endotoxin contamination must be assessed.
LPS-inducible responses in severe combined immunodeficiency (SCID) mice
Authors: Falk L.A. , McNally R. , Perera P.Y. , Kenny J. , Vogel S.N.
Published: January 2008

Lipopolysaccharide (LPS) is a potent bacterial product that has been shown to act on many different cell types both in vivo and in vitro. Injection of immunologically competent mice with LPS results in increased serum cytokine levels, followed by an array of pathophysiologic alterations that can ultimately lead to death. In this study, we examined the response of severe combined immunodeficient (SCID) mice to LPS. These mice lack mature T and B cells and have been shown to be an important model for analyzing the contribution of innate immune responses to infectious agents. Injection of SCID mice with LPS resulted in increases in CSF, TNF, and IFN levels in serum that were similar to the responses of immunocompetent controls. In response to LPS, both SCID and control mice exhibited similar levels of hypoglycemia. LPS-induced toxicity was assessed in D(+)-galactosamine-sensitized animals. SCID mice were comparably sensitive to the lethal effects of LPS as control BALB/c mice. To assess the role of natural killer (NK) cells in LPS-induced cytokine responses, BALB/c and SCID mice were injected with anti-asialo-GM1 antibody prior to injection of LPS. No significant effect on LPS-induced CSF or blood glucose levels were seen, although NK-depleted SCID mice produced somewhat more IFN in response to LPS than normal mice. Thus, NK cells are not a major source of these early LPS-induced cytokines. These data suggest that mature T and B cells and NK cells do not contribute to the initial wave of cytokines produced in response to LPS, but may contribute as secondary producers of cytokines involved in the cytokine cascade elicited by LPS injection.
Do endotoxin aggregates intercalate into phospholipid membranes in a nonspecific, hydrophobic manner?
Authors: Schromm A.B. , Brandenburg K. , Rietschel E.Th. , Seydel U.
Published: January 2008

The interaction of endotoxin aggregates with phospholipid liposomes of different composition was investigated applying fluorescence polarization spectroscopy with the fluorophore diphenylhexatriene and the resonance energy transfer technique using N-(7-nitro-2,1,3-benzoxadiazol-4-yl)-PE and N-(Rhodamine B sulfonyl)-PE. Fluorescence polarization data at constant temperature could be interpreted in favor of an intercalation of lipopolysaccharide into phospholipid liposomes even in the absence of Ca2+. Intercalation, however, could be clearly excluded from determinations performed as a function of temperature. Experiments employing the resonance energy transfer technique clearly showed that a nonspecific, hydrophobic intercalation of endotoxin aggregates into phospholipid liposomes only takes place in the presence of excess molar concentrations of divalent cations and/or after long-term incubation at elevated temperature (37°C). These findings indicate that under (near) physiological conditions nonspecific intercalation of aggregated lipopolysaccharide into phospholipid membranes represents an unlikely event. The significance of these results for an understanding of the fundamental mechanisms of cell activation by endotoxin is discussed.
Antibody cross-reactivities between gangliosides and lipopolysaccharides of Campylobacter jejuni serotypes associated with Guillain-Barré syndrome
Authors: Schwerer B. , Neisser A. , Polt R.J. , Bernheimer H. , Moran A.P.
Published: January 2008

Ganglioside-antibodies produced subsequent to Campylobacter jejuni infection may play a role in the pathogenesis of the neurological disorder Guillain-Barré syndrome (GBS). Since lipopolysaccharides (LPS) of certain C. jejuni serotypes associated with GBS (O:2, O:4, O:19) exhibit structural mimicry of gangliosides in their core oligosaccharides, we investigated antibody and ligand cross-reactivities between gangliosides and LPS of these C. jejuni serotypes. GM1-antibody reacted with O:19 LPS reflecting GM1 mimicry by the O:19 core oligosaccharide. On the other hand, asialoGM1-antibody bound to O:2 and O:19 LPS indicating a shared epitope not dependent on ganglioside mimicry. Serum IgA from GBS patients after C. jejuni infection reacted with gangliosides, predominantly GM1, and LPS of all three serotypes. Cholera toxin (GM1 ligand) recognized O:4 and O:19 LPS, whereas peanut agglutinin (Galβ1-3GalNAc ligand) recognized LPS of all three serotypes, thereby confirming structural mimicry. These results suggest that LPS from certain C. jejuni strains may function as cross-reactive antigens for anti-ganglioside B cells.
Differential susceptibility of rhesus monkeys to high doses of endotoxin
Authors: Veloso D. , Denny S. , Cosgriff T.M. , Hochstein H.D.
Published: January 2008

We investigated susceptibility of rhesus monkeys ( Macaca mulatta) to Escherichia coli endotoxin (ETX) in two ways. We infused 8 monkeys (group A) with various doses of ETX (1.0-7.5 mg/kg) to assess the effect of dose on shock severity; and we infused 6 monkeys (group B) with 1.0 mg ETX/kg to test biological variability to ETX challenge. Controls were 7 saline-infused monkeys. Systolic pressure, heart rate (HR), temperature, plasma ETX and inflammatory markers — tumor necrosis factor-α (TNF), interleukin-1 (IL-1) and IL-6 — were quantified before and at 1.5, 2.5, 6 and 26 h after infusion. The highest plasma concentrations of ETX (at 1.5 h) — < 8% that infused — correlated well with the infused doses. ETX elicited hypotension and increases in HR in all monkeys. Fever did not occur. The degree of hypotension and increase in HR and death did not correlate with ETX dose (or plasma ETX concentrations). The response of inflammatory cytokines to ETX was greater in nonsurvivors than in survivors. The observed low mortality rate (4/14) suggests that rhesus monkeys are rather resistant to high endotoxin concentrations similar to baboons but unlike humans or chimpanzees. The lack of correlation between ETX dose and shock severity suggests that there is a critical ETX concentration in each animal that leads to controllable or uncontrollable cytokine elevation in plasma, with reversible or irreversible shock, and resulting survival or death.
Effect of endotoxin on cardiopulmonary function and biosynthesis of eicosanoids and TNFα in calves chronically depleted of copper
Authors: Olson N.C. , Gengelbach G.P. , Spears J.W. , Johnson L.W. , Hellyer P.W. , Dodam J.R. , Brown T.T.
Published: January 2008

We evaluated the acute cardiopulmonary and biochemical (i.e. plasma concentrations of eicosanoids and serum levels of TNFα) response to an infusion of Escherichia coli endotoxin (8 μg/kg i.v.) in cattle that were adequate or deficient in liver and plasma copper (Cu) concentrations. Young calves were chronically fed diets containing normal Cu (control; n = 4) and low Cu (n = 14). After ~6 months of dietary treatment, the calves were anesthetized and evaluated from 0-4 h during endotoxemia. At 0 h, plasma Cu concentrations averaged 32.4% (P < 0.05) of the control value (i.e. 0.81 ± 0.08 μg/ml). Endotoxin caused a transient, early (at 20.4 min) increase in mean pulmonary arterial pressure to 114% and 147% of baseline in normal and Cu depleted calves, respectively (P < 0.05). In Cu depleted calves, endotoxin significantly decreased heart rate (0.5-1 h) and cardiac index (0.5 h) as compared to normal calves. Endotoxin caused systemic hypotension and increased TXB2 production (at 0.5 h) and these changes were potentiated in Cu depleted calves. However, after 1 h there were no statistically significant differences between the two groups of calves for any cardiopulmonary parameter evaluated nor in plasma concentrations of eicosanoids and serum levels of TNFα. We conclude that chronic dietary depletion of Cu potentiates TXB 2 biosynthesis and worsens the acute (≤1 h) cardiovascular response to endotoxin challenge in cattle.
In vitro effect of endotoxin on lipolysis and lipoprotein lipase activity in adipocytes from lean, obese and obese diabetic Zucker rats
Authors: Lanza-Jacoby S. , Rose G.L. , Rosato E.F. , Sedkova N. , Considine R.V.
Published: January 2008

Endotoxemia and sepsis in non diabetic rats are associated with alterations in adipose tissue metabolism evidenced by an increase in lipolysis and decrease in lipoprotein lipase (LPL) activity. The purpose of this study was to determine the in vitro effect of endotoxin (LPS) on lipolysis and the activities of LPL in epididymal adipocytes isolated from 11-12-week-old obese diabetic, obese, and lean rats. Epinephrine-stimulated lipolysis was higher in the adipocytes from obese diabetic and obese rats than lean rats. Maximal lipolytic response for all groups occurred with 10-5 M of epinephrine. LPS increased the lipolytic rate in adipocytes from the obese and obese diabetic rats by 58% and 97%, respectively, in comparison to the lean rats. Heparin-releasable and extractable LPL activities were suppressed in LPS-treated adipocytes from lean rats; heparin-releasable, but not extractable LPL activity, was depressed in adipocytes from obese rats. The LPS-induced depression in LPL activities did not occur in adipocytes from obese diabetic rats. Since LPL is not altered, adipose tissue from the obese diabetic rats may not become depleted when challenged with endotoxin as is the case for the normal endotoxin-treated lean rat with diminished LPL activity.
Specificity of monoclonal antibodies against Escherichia coli K-12 lipopolysaccharide
Authors: Brade Lore , Grimmecke Hans-Dieter , Holst Otto , Brabetz Werner , Zamojski Alexander , Brade Helmut
Published: January 2008

Three monoclonal antibodies against the lipopolysaccharide (LPS) of Escherichia coli strain K-12 were obtained after immunization of BALB/c mice with heat-killed bacteria and were serologically characterized by hemagglutination, enzyme-immunoassay, Western, dot, and colony blot using as antigens bacteria, LPS, de-O-acylated LPS, dephosphorylated LPS, the core oligosaccharide and synthetic partial structures representing a di-, tri-and tetrasaccharide of the K-12 outer core region. In addition, deacylated LPS and synthetic oligosaccharides were covalently linked to bovine serum albumin resulting in artificial glycoconjugate antigens. In all assays used, the antibodies were specific for K-12 LPS. These antibodies detect E. coli K-12 LPS in its isolated form as well as on the surface of bacteria and are, thus, useful for all working with this microorganism.
An N-terminal fragment of bactericidal/permeability-increasing protein protects against hemodynamic and metabolic derangements in rat Gram-negative sepsis
Authors: Steve Ammons W. , Mallari Cornell
Published: January 2008

Recombinant N-terminal fragments of bactericidal/permeability-increasing protein (BPI) are protective in acute models of endotoxemia or bacteremia. However, their usefulness in models utilizing slow infusion of bacteria has not been reported. Anesthetized rats were infused with Escherichia coli 07:K1 bacteria (5 x 109 cfu/h) for 2 h. Hemodynamics, glucose and lactate levels, tumor necrosis factor-α (TNFα) and endotoxin levels were measured for 210 min. The rats were treated with a 2 h infusion of RBPI21 or thaumatin (10 mg/kg/h), a protein that does not bind endotoxin but is similar to rBPI21 in molecular weight and isoelectric point. Control rats received only saline and vehicle. In animals treated with thaumatin, blood pressure, cardiac index and stroke volume declined rapidly (within 30 min). After 90 min glucose levels were significantly depressed whereas lactate was elevated. Endotoxemia (764 ± 353 ng/ml) was observed within 30 min. rBPI 21 significantly reduced the cardiovascular depression observed after 30 min and abolished all hemodynamic responses by the end of the experiment. rBPI21 significantly reduced hypoglycemia, elevated lactate levels, and endotoxin levels after 90 min. These results indicate that reduction of endotoxin levels resulting from RBPI21 therapy leads to significant protection in this model of acute Gram-negative bacterial sepsis.
Disparities in efficacies of anti-LPS agents in burned mouse models of LPS intoxication versus Gram-negative sepsis
Authors: Neely Alice N. , Orloff Mary M. , Holder Ian Alan
Published: January 2008

Results of anti-LPS treatments in LPS-intoxicated animals have been promising, but use of anti-LPS agents in clinical trials of sepsis have been disappointing. The purpose of this study was to determine if treatment results in intoxicated animals would hold in animal models of sepsis. Burned (15% body surface) mice were injected immediately postburn with either LPS (= intoxication) or with bacteria, which lead to sepsis. When possible, bacteria used were of the same genus and species as bacteria from which LPS was extracted for intoxication studies. Polymyxin, monophosphoryl lipid A or monoclonal antibody E5 given 1 h postburn, each protected long term against death from endotoxin intoxication. However, in septic mice, monophosphoryl lipid A and monoclonal antibody E5 did not improve survival, and polymyxin showed only transient improvement. Hence, animal models of LPS intoxication may not be optimal for testing anti-sepsis agents; use of such models, rather than of models of sepsis, may account partly for the overly optimistic expectations for anti-sepsis agents which have recently been shown to be rather ineffective in clinical trials.
Synthesis of neoglycoproteins containing 5-O-phosphorylated Kdomonosaccharide, 4-O- and 5-O-phosphorylated α-Kd↬(2 6)-2-acetamido-2-deoxy- -D-glucopyranosyl disaccharide residues
Authors: Sekljic Harald , Kosma Paul , Bartek Johannes , Fukase Koichi , Kusumoto Shoichi , Brade Helmut
Published: January 2008

Radical addition of cysteamine to anomeric allyl glycosides of Kdo 5-phosphates 1 and 4 afforded good yields of the corresponding 3-(2-aminoethylthio)propyl glycosides. Similar reactions with the α-allyl glycoside of Kdo 4-phosphate 7 led to substantial formation of the dephosphorylated product 10 through intramolecular hydrolysis of the 4-O-phosphomonoester by the terminal amino function of the spacer group. Similar side reactions occurred upon deblocking of the 4- O-phosphorylated 6-aminohexyl-α-glycoside lactone derivative 18. Allyl glycosides of 4'- O- and 5'- O-phosphorylated Kd↬ (2 6)-glucosamine derivatives were prepared in good yields via phosphorylation using the amidite procedure. The 4'-phosphate moiety was introduced following intramolecular protection of the 5'-OH group as a lactone. Subsequent deprotection gave the allyl glycosides 25 and 33 which were transformed into the corresponding stable spacer derivatives 26 and 34 by coupling with cysteamine. The ligands were activated with thiophosgene and reacted with bovine serum albumin to give the neoglycoconjugates 3, 6, 27 and 35 to be used in immunochemical studies of monoclonal antibodies directed against Haemophilus influenzae Re-mutant and related lipopolysaccharides.
Differences in mode of action of (β-lactam antibiotics influence morphology, LPS release and in vivo antibiotic efficacy
Authors: Jackson Jesse J. , Kropp Helmut
Published: January 2008

Antibiotic mediated release of endotoxin (lipopolysaccharide, LPS) from Gram-negative bacteria is implicated in septic shock. The β-lactam (cell wall active) class, in particular, has been deemed responsible for release of greater quantities of LPS than other classes of antibiotics. However, it is becoming increasingly recognized that variations in the ability to liberate free LPS exist within the β-lactam subclasses. Until recently, LPS-release studies have primarily included the cephalosporin, monobactam, and penicillin β-lactams, but not the carbapenem subclass. We document here that carbapenems significantly liberate less LPS than other β-lactam subclasses, and that disparity in LPS release also occurs within the carbapenems as well (i.e. imipenem vs meropenem). The propensity to release LPS correlates with the cumulative PBP binding affinity of each β-lactam antibiotic regardless of subclass. A direct correlation has been established between antibiotic concentration, frequency of antibiotic exposure, differential LPS release, PBP specificity (morphology, rate of bacterial lysis or cell mass) and antibiotic efficacy in two mouse models of infection. In the first, differences in the pathogenic (endotoxic) potential of potently biologically active smooth LPS (S-LPS) and poorly biologically active rough chemotype LPS (R-LPS) expressing bacteria underscores the in vivo importance of endotoxin in mediating lethality following antibiotic chemotherapy. Endotoxin is, however, not innately toxic and thus its lethal effects are mediated through LPS stimulation of host cells (i.e. macrophages) to produce deleterious levels of cytokines. In the second experimental model, therefore, the toxic effects of antibiotic-liberated endotoxin have been abrogated by pretreatment of mice with carrageenan (CGN) prior to challenge with endotoxin or Gram-negative bacteria. CGN treatment eliminates or markedly reduces the numbers of LPS-responsive mediator cells thus affecting mortality. In both animal models, an in vivo role for LPS is demonstrated through differential changes in antibiotic efficacy (ED 50) in response to differential effects of LPS via modulation of the endotoxic sensitivity/responsiveness of the host. We conclude that antibiotic class, concentration, dosing intervals (timing) and perhaps, method of treatment (i.e. bolus vs infusion) may each be important in the survival of experimental animals severely infected with Gram-negative bacteria.