bluuebunny/crossref_metadata_2025 huggingface dataset:
Regional Policy Differences In the UK: Education in Wales
Authors: Farrell Catherine M. , Law Jennifer
Published: January 2008

This paper is concerned with regional policy differences in the UK. The classification of education in the UK as a concurrent rather than an exceptional policy by Rose (1982) is evaluated in this paper. It is argued that the Rose framework does not distinguish between policy content and policy impact. In failing to distinguish between these elements of policy, Rose and other authors implythat regional policy differences within the UK are minimal. However, whilst some policies can have the same content, they may havea different impact within the separate nations of the UK. The finding of this paper is that education policy in England and Wales is broadly concurrent in its content and exceptional in its impact in Wales. This is due to the role and influence of the education policy community in Wales.
Public Services under New Labour: collaborative discourses and local networking
Authors: Clarence Emma , Painter Christopher
Published: January 2008

A by-product of the fragmentation of services that occurred at local government level under the Conservative Government was the emergence of networks and other forms of joint working. These became formalised particularly through various economic and urban regeneration schemes. But elsewhere they were often an ad hoc response to the changing local governance structures. The election of the Labour Government in 1997 has, this article contends, seen the emergence of a more collaborative discourse in which networking and partnership are the intentional outcomes of many of the central government's policy initiatives, including the Education and Health Action Zones and the Social Exclusion Unit. This collaborative discourse is founded on a perception of the importance of trusting, mutually beneficial relationships, as distinct from adversarial, low trust relationships, to achieving central government aims and objectives. Based on interviews with local authority Leaders and Chief Executives, and other 'elite' actors, the article explores how well local authorities are responding to the challenges and opportunities that these policy initiatives will bring both with regard to internal structures and decision making processes, as well as to the role of the local authority in local governance networks.
The response of GP fundholders and practice managers to the reorganisation of primary care services
Authors: Coleshill Paul , Goldie Duncan , Sheffield James W.
Published: January 2008

This article aims to answer a number of linked questions. The first involved a detailed recording of management behaviour and management skills as demonstrated within the General Practitioner Fundholding (GPFH) practice. The purpose for doing this was to determine whether or not GPFHs had acquired management skills, and whether or not these skills had been used to benefit primary care. If GPFHs demonstrated such skills, the second objective was to determine the use to which these skills would be put in the proposed reorganisation of primary care. In order to do this, we examined the proposed structure of primary care in Scotland, and asked GPFHs about the consultation process and their role in the proposed structure.
S-form LPS induces leukocyte adhesion to human vascular endothelial cells as potent as IL-1: lipid A precursor Ia antagonizes induction of adhesion by LPS
Authors: Schönbeck U. , Flad H.-D. , Rietschel E. Th. , Brandt E. , Loppnow H.
Published: January 2008

The expression of adhesion molecules on human vascular endothelial cells (EC) during infection or inflammation is necessary to initiate migration of leukocytes to the disease focus. Various bacteria or endotoxins (lipopolysaccharide, LPS) might exert differential capacity to induce adhesion molecules in endothelial cells. Therefore, we first investigated induction of adhesion by bacteria. Heat-killed Gram-negative bacteria (Escherichia coli) induced adhesion, whereas the Gram-positive Bacillus subtilis and Staphylococcus epidermidis or Mycobacterium tuberculosis did not, indicating the significance of LPS for this activation. Purified S-form LPS stimulated endothelial cells to express adhesion molecules for polymorphonuclear cells (PMN) or lymphocytes in a dose-dependent fashion. S-form LPS, containing an 0-specific chain, induced a maximal level of adhesion, comparable to adhesion induced by interleukin-1 (IL-1). We obtained the same results in cell ELISA with anti-ICAM-1 antibody (84H10). R-form LPS and free lipid A, lacking an O-specific chain also stimulated adhesion, however, to a lower degree (39-60%). Synthetic lipid A precursor Ia (compound 406) or another LPS-antagonist (non-toxic Rhodobacter capsulatus LPS) did not trigger endothelial cells to express adhesion molecules. These antagonists specifically inhibited LPS- or free lipid A-, but not IL-1-induced adhesion. These results suggest that lipid A is the active structure of LPS necessary for induction of adhesion, that the oligosaccharide portion is important for the capacity of LPS to stimulate adhesion, and that activation of adhesion to endothelial cells by LPS requires specific LPS binding sites.
Inhibition of LPS binding on human monocytes by phosphonooxyethyl analogs of lipid A
Authors: Heine H. , Brade H. , Kusumoto S. , Kusama T. , Rietschel E. Th. , Flad H.-D. , Ulmer A.J.
Published: January 2008

We investigated the inhibition of LPS binding on human monocytes by synthetic analogs of lipid A. A common characteristic of the analyzed structures is a α-(or β-) phosphonooxyethyl group in position 1 of the GlcN I of the lipid A backbone. Compounds PE-1, PE-2 and PE-3 are analogs of synthetic Escherichia coli lipid A whereas PE-4 represents an analog of tetraacyl precursor Ia (synthetic compound 406). By determining the ability of these preparations to inhibit the binding of FITC-labeled LPS (E. coli 0111:B4) on human monocytes the relationship between their structure and cellular binding affinity was evaluated. The results showed a structure-dependent hierarchy of inhibition capacity. Thus, compound PE-1 inhibited the binding of FITC-LPS only slightly more than PE-2. However, compound PE-3, possessing β-configurated GlcN I, exhibited a drastically decreased inhibition capability. Best inhibition was obtained with compound PE-4. It was furthermore shown by a Lineweaver-Burk plot that the inhibition of LPS binding was due to competition of FITC-LPS and PE-4 for the same binding structure. The synthesis of stable 1-phosphonooxyethyl analogs of precursor Ia with high affinity for LPS receptor structures but lacking cytokine-inducing capacity (like PE-4) may be of relevance for their function as potent antagonists of LPS in therapy of endotoxic shock and sepsis.
Endotoxin tolerance is not a LPS-specific phenomenon: partial mimicry with IL-1, IL-10 and TGFβ
Authors: Cavaillon J.-M. , Pitton C. , Fitting C.
Published: January 2008

Administration of non-lethal doses of lipopolysaccharide (LPS) to experimental animals and humans results for a short period of time in a state of hyporesponsiveness to a second LPS challenge. This phenomenon, known as endotoxin tolerance, has been reproduced in vitro using human monocytes, rendered endotoxin-tolerant following a first incubation with LPS. A further activation by LPS was manifested by a dramatically reduced production of tumor necrosis factor α (TNFα). We report this low responsiveness of LPS pretreated monocytes as an endotoxin non-specific phenomenon. Indeed, TNFα release upon further activation with either killed Gram-positive bacteria (Staphylococci, Streptococci) or zymosan was also significantly diminished. This was not the case when phorbol myristate acetate (PMA) was used as a second triggering agent, suggesting that the monocyte hyporesponsiveness due to LPS does not affect all activation pathways, particularly that of protein kinase C. On the other hand, both PMA and zymosan pretreatment could reduce a further activation of monocytes by LPS. We investigated whether the first signal(s) delivered by LPS, could be mimicked by some of the LPS-induced cytokines. Preincubation of monocytes with either interleukin-1 (IL-1), IL-10 or transforming growth factor β (TGFβ) lower the TNFα production upon further activation with LPS. None of the cytokines alone was as efficient as the LPS molecule, but high levels of tolerization were obtained with combination of IL-1, IL-10 and TGFβ. Neither IL-6, IL-8 nor TNFα led to hyporeactive cells. Our data indicate that endotoxin tolerance is not an LPS-specific phenomenon and that more than one cytokine can contribute to render human monocytes hyporeactive to further activation by LPS.
Structural analysis of the lipid A derived from the lipopolysaccharide of Brucella abortus
Authors: Qureshi N. , Takayama K. , Seydel U. , Wang R. , Cotter R.J. , Agrawal P.K. , Bush C.A. , Kurtz R. , Berman D.T.
Published: January 2008

Lipopolysaccharide (LPS) of Brucella abortus strain 45/20 was purified using a novel method. Monophosphoryl lipid A (MPLA) was prepared from this LPS, methylated, and purified by high performance liquid chromatography. Chemical, mass spectral, and nuclear magnetic resonance analyses showed that MPLA consists of heptaacyl lipid As with molecular weights of 2095, 2123, 2151 and 2179. They contained the β-1,6-linked 2,3-diamino-2,3,-dideoxy-glucose disaccharide backbone and a phosphate group at the 4' position. Bisphosphoryl lipid A was also prepared and completely O-deacylated. It contained an additional phosphate group, and either 2 hydroxyhexadecanoic, 1 hydroxytetradecanoic, 1 hydroxydodecanoic acids or 2 hydroxyhexadecanoic and 2 hydroxydodecanoic acids, all in amide linkage. The predominant ester-linked fatty acyl group in acyloxyacyl linkage was hexadecanoate.

The purified LPS, bisphosphoryl lipid A, and MPLA from B. abortus showed about 14%, 3% and 1%, respectively, of the B cell mitogen activity of ReLPS from Escherichia coli at 1.0 μg/ml.
Characterization and sequence analysis of the lsg (LOS synthesis genes) locus from Haemophilus influenzae type b
Authors: McLaughlin R. , Lee N.-G. , Abu Kwaik Y. , Spinola S.M. , Apicella M.A.
Published: January 2008

Analysis of the lsg (LOS synthesis genes) cluster in Escherichia coli strain K12 and mutations in the lsg locus in Haemophilus influenzae type b indicated the presence of 3 regions responsible for sequential modifications of E. coli lipopolysaccharide (LPS). Sequencing of the lsg region yielded 7,435 bp that encompassed 7 complete and 1 partial open reading frames (ORFs 1-8). The predicted product of ORF1 had homology to the consensus sequence of cytochrome b proteins (21% identity, 51% similarity) and to other transmembrane proteins. The products of ORF5 and ORF6 share overall 23% identity and 49% similarity with each other. The ORF6 protein had high homology with the product of ORF275 of the E. coli rfb gene cluster (40% identity, 58% similarity), whose function is not known. Multiple sequence alignment of the ORF5 and ORF6 proteins with the RfbB, RfbJ and RfbX proteins revealed conserved motifs over the N-terminal half region of all these proteins. The products of ORF7 and ORF8 are homologous with Azotobacter vinelandii MolA protein (30% identity, 51% similarity) and MolB protein (26% identity, 48% similarity), respectively. The promoter regions of ORF1, 7 and 8 were determined by primer extension analysis and found to be similar to bacterial σ70-dependent promoters. ORF7 and ORF8 are transcribed into diverse orientation. At least 5 of the encoded proteins have been identified using coupled E. coli transcription/translation system and labeling with [35S]-methionine. We conclude that the genetic organization of the lsg biosynthesis pathway involves multiple operons that lead to the assembly of an H. influenzae LOS structure.
Deactivation of the respiratory burst of human and murine macrophages by lipopolysaccharide is a receptor mediated effect independent from autocrine mechanisms
Authors: Schneemann M. , Schaffner A.
Published: January 2008

Endotoxin blunts secretion of reactive oxygen intermediates by macrophages. Because transforming growth factor β (TGFβ) or interleukin 10 (IL-10) were shown to deactivate macrophages, and are secreted by macrophages in response to LPS, we sought for autocrine mechanisms of macrophage deactivation by endotoxin. TGFβ did not deactivate the respiratory burst of human blood-derived and resident or thioglycollate-induced murine peritoneal macrophages. According to previous reports, suppression of H202 secretion by TGFβ was restricted to periodate-elicited murine peritoneal macrophages.

In contrast to TGFβ, IL-10 deactivated systems producing reactive oxygen intermediates in human blood-derived human macrophages, but neither anti-TGFβ1 nor anti-IL-10 antibodies restored LPS-mediated deactivation of macrophages.

Supernatants from LPS-treated human blood-derived macrophages could not confer deactivation to homologous macrophages.

Deactivation by LPS required the presence of serum proteins and appeared to be mediated by the CD14 antigen-related LPS receptor, because anti-CD14 antibody and IL-4, which downregulates CD14, antagonized the LPS effect.

Taken together, these observations indicate that the suppressive effect of LPS on the respiratory burst of macrophages is lipid A receptor-dependent, and results in a direct deactivation response of the cell without mediation by autocrine mechanisms.
Contribution of clonally distinct subpopulations to heterogeneous production of inducible nitric oxide synthase by LPS-stimulated mouse macrophages
Authors: Alley E.W. , Zhang X. , Russell S.W.
Published: January 2008

Expression of the inducible nitric oxide synthase (iNOS) gene in mouse macrophages can be induced by bacterial lipopolysaccharide (LPS). iNOS (EC 1.14.13.39) catalyzes production of the reactive nitrogen intermediate, nitric oxide (NO), which is very important to macrophage-mediated host defense in species such as the mouse and rat. We have investigated production of iNOS at the level of single cells through immunocytochemical analysis of LPS-stimulated macrophages. Both bone marrow culture-derived macrophages and those of the cell line, RAW 264.7, were examined. Heterogeneous production of iNOS within macrophage populations was explained in part by the existence of clones that were high producers of iNOS and, therefore, NO, while other clones were reproducibly low producers of each. All clonally derived populations continued to demonstrate heterogeneous iNOS production, suggesting that at least one additional mechanism must be responsible for the phenomenon of heterogeneity. In contrast to iNOS, LPS-stimulated macrophages synthesized interferonβ (IFNβ) uniformly throughout the population.
Synthesis of Kdo-trisaccharide derivatives of chlamydial and enterobacterial LPS containing carboxyl-reduced or β-configurated Kdo-residues
Authors: Kosma P. , D'Souza F.W. , Brade H.
Published: January 2008

Five allyl glycosides corresponding to the 3-deoxy-D- manno-2-octulosonic acid (Kdo) containing genus-specific LPS epitope of Chlamydia were synthesized. Compounds 5 and 22 contain one carboxyl-reduced Kdo moiety linked to O-4 of the proximal Kdo unit, whereas the analogues 28, 34 and 36 each contain one β-linked Kdo-residue within the trisaccharide sequence Kdo p-(2→8)-Kdo p-(2→4)-Kdo p. Elaboration of the carboxyl-reduced derivatives was achieved by BF3•Et2O-catalyzed coupling of Kdo-fluoride derivatives 1 or 14 with the 7,8- O-carbonyl-derivative 2. The β-linked oligosaccharides were obtained by Helferich-glycosidation of the respective Kdo-disaccharide bromide derivatives 26 and 31. The deprotected compounds - characterized by H and 13C NMR spectroscopy - are suitable haptens for the immunochemical study of monoclonal antibodies directed against the Kdo-region of chlamydial and enterobacterial LPS.
Characterization of the interaction of lipid A and lipopolysaccharide with human serum albumin: implications for an endotoxin carrier function for albumin
Authors: David S.A. , Balaram P. , Mathan V.I.
Published: January 2008

The interactions of lipid A and lipopolysaccharide (LPS) with human serum albumin (HSA) were examined using fluorescence methods. Lipid A binds HSA with a stoichiometry of 2:1 with dissociation constants of 1.0 μM and 6.0 μM for the high- and low-affinity interactions, respectively. Lipid A displaces HSA-bound dansylsarcosine competitively, but not HSA-bound warfarin, suggesting that domain III-A, and not domain 11-A, is a lipid A binding site. Domain I does not contribute a site for lipid A. Based on these data, and the structural similarity between subdomains III-A and III-B, it is proposed that these two regions of HSA represent the high- and low-affinity sites of interaction of lipid A. Whole LPS also binds HSA, displacing dansylsarcosine, and its lipid A moiety appears to be the interaction site. However, there are differences between LPS and free lipid A. Polymyxin B forms ternary complexes with LPS bound to HSA, suggesting that the regions on LPS recognized by HSA and polymyxin B are different. The observed affinity of lipid A for HSA and mass action effects due to its abundance in the circulation would imply a major LPS carrier function for HSA.
A synthetic lipid A analog, B464, provides significant protection against the cardiopulmonary derangements in porcine Gram-negative sepsis
Authors: Ridings P. , Windsor A.C. , Rossignol D.P. , Christ W.J. , Blocher C. , Fisher B.J. , Fowler A.A. , Sugerman H.J.
Published: January 2008

Many of the cardiopulmonary derangements associated with Gram-negative sepsis result from the activation of monocytes and macrophages by endotoxin with the resultant release of humoral mediators such as TNFα. Lipid A has been shown to retain the majority of endotoxin toxicity, however lipid A from nontoxic organisms has been shown, in vitro, to antagonize the action of other toxic endotoxin species. We examined the effects of a synthetic analog of lipid A (B464), on the evolution of hemodynamic derangements and acute lung injury following experimental Gram-negative sepsis. Anesthetized, ventilated swine were made septic with a 1 h intravenous infusion of live Pseudomonas aeruginosa and studied for 5 h. A treatment group received a bolus of B464 (100 μg/kg) prior to sepsis and a 1 h infusion (100 μg/kg/h) during the P. aeruginosa induction. A control group received simply 0.9% saline. All animals were studied for 5 h. B464 treatment failed to alter septic pulmonary hypertension or the decline in cardiac output. Late recovery from systemic hypotension was associated with reversal of arterial acidosis. Septic neutropenia was unaltered and was associated with increased lung neutrophil (PMN) sequestration measured by lung myeloperoxidase activity. However, decreased bronchoalveolar lavage protein content and improved arterial oxygen tension indicated attenuated acute lung injury in B464 treated animals. These preliminary data indicate that B464 may prove to be an effective tool in the treatment of human sepsis.
G-CSF - an anti-inflammatory cytokine
Authors: Hartung T. , Volk H-D. , Wendel A.
Published: January 2008

We have previously reported that in various macrophage populations prepared from G-CSF treated rats LPS-inducible TNF release was suppressed. In vitro, LPS induced liver cell death only when hepatocytes were cocultured with liver macrophages. Rat Kupffer cells from G-CSF treated donor animals were less potent in mediating LPS-inducible hepatocytotoxicity in vitro than cells from control animals. These ex vivo findings were confirmed in vivo by demonstrating that G-CSF treatment attenuated LPS-inducible circulating TNF levels and protected from liver injury and mortality.

We extended these observations to humans in two studies with G-CSF treated volunteers. In a pilot study, 11 subjects were treated single-blindly with 480 μg G-CSF s.c. (n = 7) or saline placebo (n = 4). Blood was taken at different time-points relative to G-CSF injection and cytokine release capacity was assessed in LPS stimulated whole blood incubations. In blood from G-CSF treated volunteers, we found reduced LPS-inducible TNF formation while the release of both soluble TNF receptor (sTNF-R) and interleukin 1 receptor antagonist (IL-1ra) were increased. In a second double-blind, randomized and controlled study, three groups of seven volunteers were treated once or twice 24 h apart with G-CSF or solvent placebo. Besides LPS, various stimuli were included to initiate cytokine release in a whole blood assay. The reduction of TNF formation (mean 53 % at 24 h after G-CSF) was different with the various stimuli. All stimuli increased IL-1ra (mean 14-fold) and sTNF-R (mean 3-fold) at 24 h after G-CSF. LPS-inducible IFN-γ and GM-CSF were significantly reduced. Our data indicate that the pattern of cytokines produced by human whole blood taken after G-CSF treatment in response to a variety of stimuli is shifted from pro- to anti-inflammatory mediators. These findings extend the knowledge on the pharmacology of G-CSF in animal models of the systemic inflammatory response syndrome and prompt a trial of G-CSF prophylaxis with this indication.
Lipopolysaccharide induces expression of Egr-1 in macrophages through upstream CArG box (serum response) elements
Authors: Li C. , Mitchell D.H. , Coleman D.L.
Published: January 2008

The protein product of the early response gene, Egr-1, is required for macrophage differentiation. Egr-1 mRNA is induced by lipopolysaccharide within minutes in murine peritoneal macrophages through an effect on the rate of transcription of Egr-1. Therefore, we undertook a series of studies to characterize Egr-1 protein induced by LPS and to define enhancer elements in the Egr-1 promoter which mediate the effect of LPS in murine macrophages. Salmonella minnesota Re 595 LPS (10 ng/ml or higher) induced Egr-1 protein of 80 kDa within 45 min in thioglycollate-elicited peritoneal macrophages. The promoter region of Egr-1 contains six serum response elements (SRE) which each contain a core CArG box. CAT activity in LPStreated macrophages transfected with Egr-1-CAT constructs containing the two most upstream 5' SREs was increased 4.7-fold whereas CAT activity in macrophages transfected with constructs lacking these two most upstream SREs was not affected by LPS. LPS induced CAT activity in macrophages transfected with constructs containing tetramers of each of the two most 5' SREs upstream of a heterologous promoter linked to CAT. Mutation of the CArG box abolished the effect of LPS. These findings suggest that the CArG box elements within the two most upstream SREs may mediate the effect of LPS on transcription of Egr-1.
Interactions of linear dicationic molecules with lipid A: structural requisites for optimal binding affinity
Authors: David S.A. , Mathan V.I. , Balaram P.
Published: January 2008

The structural determinants of the binding affinity of linear dicationic molecules toward lipid A have been examined with respect to the distance between the terminal cationic functions, the basicity, and the type of cationic moieties using a series of spermidine derivatives and pentamidine analogs by fluorescence spectroscopic methods. The presence of two terminal cationic groups corresponds to enhanced affinity. A distinct sigmoidal relationship between the intercationic distance and affinity was observed with a sharp increase at 11 Å, levelling off at about 13 Å. The basicity (pK) and nature of the cationic functions are poor correlates of binding potency, since molecules bearing primary amino, imidazolino, or guanido termini are equipotent. The interaction of pentamidine, a bisamidine drug, with lipid A, characterized in considerable detail employing the putative intermolecular excimerization of the drug, suggests a stoichiometry of 1:1 in the resultant complex. The binding is driven almost exclusively by electrostatic forces, and is dependent on the ionization states of both lipid A and the drug. Under conditions when lipid A is highly disaggregated, pentamidine binds specifically to bis-phosphoryl- but not to monophosphoryl-lipid A indicating that both phosphate groups of lipid A are necessary for electrostatic interactions by the terminal amidininium groups of the drug. Based on these data, a structural model is proposed for the pentamidine-lipid A complex, which may be of value in designing endotoxin antagonists from first principles.
A comparative study of biological activities of lipoteichoic acid and lipopolysaccharide
Authors: Renzi P.M. , Lee C-H.
Published: January 2008

Lipoteichoic acids (LTAs) of Gram-positive bacteria and lipopolysaccharide (LPS) are structurally similar and share several biological activities. In this study, the capacity of LTAs for inducing the cell surface expression of adhesion molecules ICAM-1, VCAM-1, and ELAM-1 by HUVEC were investigated and compared to that of LPS. In the presence of serum, LTA from Staphylococcus aureus, Streptococcus sanguis, Streptococcus pyogenes and Streptococcus mutans dose-dependently stimulated adhesion molecule expression above control levels although they were 100-1000 times less potent than LPS. LTA from Streptococcus faecalis and Bacillus subtilis had no effects. Similar to LPS, the activity of the LTAs was dose-dependently inhibited by polymyxin B, whereas unlike LPS, LTA activity decreased in the presence of triethylamine. The LTAs also exhibited reactivity in the Limulus amebocyte lysate (LAL) assay correlating with their ability to induce ICAM-1 expression. However, their LAL reactivity was approximately 3-6 orders of magnitude less than that of LPS. Furthermore, LTA-stimulated ICAM-1 expression was serum-dependent and inhibited by anti-CD14 antibody, 63D3. Thus, LTA shares several properties with LPS including inhibition by polymyxin B, reactivity in the LAL assay, and stimulation of endothelial cells via a CD14-dependent mechanism. These results suggests a possible role for LTAs in the inflammatory response due to infections with Gram-positive bacteria.
Inhibition of nitric oxide formation does not affect endotoxin lethality in rats
Authors: Zurovsky Y. , Eligal Z.
Published: January 2008

Endotoxin causes release of nitric oxide (NO) from different types of cells. A causative role has been suggested for NO in endotoxin-induced hypotension. In order to test the hypothesis that NO may be involved in the high mortality rate following endotoxin challenge, the competitive NO synthase inhibitors Nw nitro-L-arginine methyl ester (L-NAME) and NG methyl L-arginine (L-NMA), or the NOS substrate L-arginine, were administered to rats in the presence and absence of endotoxin. Death rates were examined after drug injection. Urine nitrite concentration and mean arterial blood pressure were examined for 24 h following endotoxin injection. It was found that endotoxin injection increased mortality rates. The mortality rate of rats injected with NOS inhibitors or an NOS substrate concomitantly with endotoxin was similar to that observed in rats treated with endotoxin alone. Endotoxin elicited a decrease in mean arterial blood pressure, whereas treatment with NOS inhibitors prevented this decrease. Treatment with L-arginine markedly exacerbated the endotoxin effects on blood pressure and resulted in hypotension 6 h after the endotoxin injection. The urine nitrite concentration increased after injection of endotoxin or L-arginine alone, or after injection of endotoxin concomitantly with L-arginine. The urine nitrite concentration after injection of endotoxin together with L-NAME was smaller than after injection of endotoxin alone. The effects of L-NAME on the urine nitrite concentration and on the blood pressure were reversed with L-arginine. These results do not support the hypothesis that NO mediates the high mortality rates following administration of endotoxin.
In vivo neutralization of tumor necrosis factor attenuates the generalized Shwartzman reaction in the rabbit
Authors: Ueda Yoshihiko , Kwang Hoon Lee  , Ito Shogo , Saito Kenichi , Niesen Neal , Brentjens Jan R. , Gans Reinold O.B.
Published: January 2008

A role for tumor necrosis factor (TNF) and TNF-induced tissue factor (TF) expression has been postulated in endotoxin (LPS)-induced diffuse intravascular coagulation (DIC). We studied the protective efficacy of goat polyclonal gammaglobulin (lgG) against recombinant human (rh)-TNF in the generalized Shwartzman reaction (GSR) in rabbits, a model of DIC. Administration of anti-rhTNF 2 h before the first, preparative, LPS injection afforded 95% neutralization of TNF released during the GSR. Compared to saline-pretreated controls, a modest attenuation of fibrin deposition in kidney, lung, liver and spleen was observed 4 h after the second provocative LPS injection. Except for the lung, fibrin deposition in the anti-rhTNF group was not attenuated compared to the non-immune goat IgG (ni-lgG)-pretreated group. Both ni-lgG and anti-rhTNF prevented LPS-induced TF expression by mononuclear cells, but not by isolated glomeruli. Other mediators than TNF and TF are involved as well in glomerular injury and fibrin deposition after LPS.
Antibiotic induced release of endotoxin - clinical data and human studies
Authors: Prins Jan M.
Published: January 2008

In vitro and animal studies, but also clinical data, indicate that treatment with antibiotics may cause systemic or local endotoxin liberation. In several studies this so-called 'endotoxin release' was accompanied by a deterioration of parameters of illness. In addition, in vitro studies have clearly demonstrated that equally effective antibiotics can differ in the amount of endotoxin they release. These differences between antibiotics can be related to different modes of anti-bacterial activities. In a number of animal studies these differences in endotoxin-releasing potential resulted in differences in morbidity and/or mortality. In only one study, antibiotics were compared in this respect in septic patients. The results of that study suggested that differences between antibiotics in endotoxin release may indeed affect the inflammatory response during treatment.
Resistance of Serratia marcescens to polymyxin B: a comparative investigation of two S-form lipopolysaccharides obtained from a sensitive and a resistant variant of strain 111
Authors: Seltmann G. , Lindner B. , Holst O.
Published: January 2008

Lipopolysaccharide (LPS) of the polymyxin B (PmB)-sensitive variant of Serratia marcescens smooth-type strain 111 (senLPS) binds significantly more PmB than that of the resistant variant (resLPS), following mild acid hydrolysis this difference is abolished. The main compositional differences between these strains are: (i) the amount of 4-amino-4-deoxy-arabinose (Ara4N); and (ii) the substitution patterns of the phosphate groups. ResLPS contains three times more Ara4N than senLPS, mainly phosphodiesters, whereas senLPS contains mainly (di)-phosphomonoesters. Thus, the senLPS, and the bacterial surface, contain a higher negative charge density. Mild acid hydrolysis decreases the PmB-binding capacity of senLPS and increases that of resLPS. The resulting change in LPS composition was a reduction in the amount of phosphate (about 40%) and a loss of nearly all the Ara4N residues, from both LPS. Mild alkaline hydrolysis removes Ara4N from both LPS, and the hydrolysate contains Ara4N-phosphate, but no Ara4N-(1 →8)-Kdo was detected. The results suggest that the 4'-phosphate of lipid A mediates the binding of PmB, whereas its substitution by Ara4N prevents it.
α-Tocopherol liposomes alleviate LPS-induced hepatotoxicities
Authors: Suntres Z.E. , Shek P.N.
Published: January 2008

Gram-negative bacteria, in part through lipopolysaccharide (LPS) and tumor necrosis factor-alpha (TNFα), activate phagocytes to generate reactive oxygen species (ROS), which have been known to play a key role in the pathogenesis of liver injury. Accordingly, we hypothesized that the susceptibility of the liver to ROS should be reduced by augmenting its antioxidant status. Adult male Sprague-Dawley rats were pretreated with α-tocopherol liposomes (20 mg α-tocopherol/kg body weight, i.v.), plain liposomes or saline. 24 h after liposomal treatment, rats were injected intravenously with LPS (1 mg/kg, Escherichia coli: 0111:B4) and killed 2 h later. Livers of saline-pretreated animals challenged with LPS were damaged as demonstrated by increases in plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities. The hepatic injury appeared to be associated with oxidative stress-mediated mechanisms as evidenced by increases in lipid peroxidation and decreases in glutathione concentration in the liver, both indices of oxidative stress. Also, LPS injection resulted in increases in plasma TNFα and thromboxane B2 (TXB2) levels, as well as increases in hepatic myeloperoxidase (MPO) activity and chloramine concentration, suggestive of activation of the inflammatory response. Pretreatment of rats with plain liposomes, 24 h prior to LPS challenge, failed to protect against the LPS-induced liver injury. Although pretreatment of animals with α-tocopherol liposomes was not effective in preventing the LPS-induced inflammatory response, it conferred a partial protection against the LPS-induced changes in plasma AST and ALT activities as well as in hepatic levels of lipid peroxidation, glutathione and chloramine concentrations. These data appear to suggest that augmentation of the hepatic antioxidant status is effective in alleviating the LPS-induced liver injury.
Review: Molecular mimicry of host structures by lipopolysaccharides of Campylobacter and Helicobacter spp.: implications in pathogenesis
Authors: Moran A.P. , Appelmelk B.J. , Aspinall G.O.
Published: January 2008

Molecular mimicry of host structures by the saccharide portion of lipopolysaccharide (LPS) contributes to the virulence of certain strains of mucosal pathogens. Mimicry by the low molecular weight (low-Mr) LPSs of Neisseria and Haemophilus spp. have been the most extensively studied. However, studies within the last decade have revealed other types of mimicry within the saccharide moieties of LPSs of the enteric pathogen Campylobacter jejuni and the gastroduodenal pathogen Helicobacter pylori. The core oligosaccharides of low-Mr LPSs of C. jejuni serotypes which are associated with the development of Guillain-Barré syndrome (GBS), a neurological disorder, exhibit mimicry of gangliosides. Cross-reactive antibodies between LPSs and gangliosides which are induced during antecedent C. jejuni infection are considered to play an important role in GBS pathogenesis. The O-polysaccharide chains of high-Mr LPSs of a number of H. pylori strains mimic Lewisx and/or Lewisy blood group antigens. This mimicry may camouflage the bacterium in the gastric mucosa upon initial infection. With the progression of infection, the mimicry may play a role in immune response regulation and the induction of autoantibodies against the gastric proton pump, a glycoprotein that also expresses Lewis antigens.
Erythrocyte-bound interleukin-8 and RANTES in human endotoxemia
Authors: Manten A. , Pajkrt D. , de Winter R.J. , de Jong Y.P. , Jansen J. , ten Cate J.W. , van Deventer S.J.
Published: January 2008

Erythrocytes have been suggested to bind chemokines, thus limiting their effective concentration in the circulation. We investigated binding of the chemokines IL-8 and RANTES to erythrocytes following endotoxin challenge. Endotoxin caused a transient increase of IL-8 in plasma, and a gradual increase of erythrocyte-bound IL-8, remaining elevated for at least 24 h in vivo. Binding of IL-8 to erythrocytes was confirmed by incubation of freshly isolated erythrocytes with endotoxin stimulated plasma. Incubation for 1 h led to a decrease in plasma concentrations of IL-8 with a concomitant increase in erythrocyte-bound IL-8 levels. Erythrocyte-bound RANTES was induced early following endotoxin infusion and our data may suggest competition between the two chemokines for the same binding site on erythrocytes in vivo. In conclusion, in humans, both IL-8 and RANTES are induced by endotoxin and are, at least in part, cleared from the plasma by binding to erythrocytes.
Inhibition of antibiotic-induced lysis and lipopolysaccharide release from Escherichia coli 0111 :B4 by egg white lysozyme
Authors: Liang A.H. , Ohno N. , Yadomae T.
Published: January 2008

Experimental and clinical data have demonstrated that antibiotics especially β-lactams, have the release of lipopolysaccarides (LPS) as adverse effects. Our previous studies showed that lysozyme (LZM) has the capacity to interact with LPS to neutralize toxicity of LPS. In this study, we examined the effect of egg white lysozyme (EW-LZM) on β-lactam-induced lysis and LPS release from Escherichia coli O111:B4. Ampicillin (ABPC) and cefotaxime (CTX) induced lysis of E. coli O111:B4 and released large amounts of LPS into culture supernatants, and thus generated excess tumor necrosis factor α (TNFα) from macrophages both in vitro and in vivo. Administration of EW-LZM in combination with ABPC or CTX (LZM concentration > 1 mg/ml) still showed bactericidal activity but without bacteriolysis. ABPC or CTX-induced LPS release as well as TNFα productivity were significantly decreased by the presence of EW-LZM. Instability of EW-LZM/β-lactam treated cells was confirmed by the fact that: (i) the cells were easily lysed by washing in the absence of EW-LZM; and (ii) a significant amount of a periplasmic enzyme, alkaline phosphatase was detected in the culture supernatant. These results suggested that interaction of LZM with LPS protects E. coli O111:B4 from β-lactam-induced lysis and LPS release without increasing viability.
Review: Evidence against the hypothesis that antibodies to the inner core of lipopolysaccharides in antisera raised by immunization with enterobacterial deep-rough mutants confer broad-spectrum protection during Gram-negative bacterial sepsis
Authors: Greisman S.E. , Johnston C.A.
Published: January 2008

Antisera to rough enterobacterial mutants of chemotypes Ra, Rc, and Re have been reported to confer broad-spectrum protection against wild-type smooth strains. It has been hypothesized that binding and neutralization of lipopolysaccharides (LPS) by antibodies to common core epitopes underlies such protection. This review summarizes experiments by our laboratory and others that do not confirm this concept and proposes reasons for the divergent results. Studies indicating broad-spectrum protection by rough-mutant antisera often had defects in experimental design or methodology. These include the failure: (i) to use matched pre- and postimmune sera from the same donors to control for variable protective activity of normal sera; (ii) to exclude the role of natural and polyclonally stimulated antibodies with proven protective activity against the infecting bacterial strain (e.g. O-specific, capsular, Pseudomonas exotoxin A); (iii) to exclude protective effects of acute-phase serum factors; (iv) to exclude protective effects of endotoxin contamination after adsorption or fractionation of antibody preparations; (v) to use non-boiled bacteria and LPS not subjected to acid-hydrolysis or gel-fractionation, and to exclude nonspecific adsorption, to demonstrate physiologically meaningful binding of rough-mutant antibodies to smooth enterobacteria and their LPS.
Protection against lethal polymicrobial sepsis by CNI-1493, an inhibitor of pro-inflammatory cytokine synthesis
Authors: Villa P. , Meazza C. , Sironi M. , Bianchi M. , Ulrich P. , Botchkina G. , Tracey K.J. , Ghezzi P.
Published: January 2008

Polymicrobial sepsis caused by cecal ligation and puncture (CLP) in mice produces the inflammatory and pathological sequelae of lung neutrophil infiltration, adult respiratory distress syndrome (ARDS) and death. These sequelae are dependent upon the synergistic interaction between several inflammatory mediators, including tumor necrosis factor (TNF), interleukin 1 (IL-1 ), and nitric oxide (NO). The overlapping spectrum of multiple mediator toxicity has hampered efforts to develop therapies for sepsis based on selective inhibition of a single mediator. Therefore, we tested the hypothesis that inhibition of multiple pro-inflammatory mediators would abrogate lethality. Our results show that administration of a tetravalent guanylhydrazone compound (CNI-1493) protected mice against 10 day mortality in CLP. Evidence of suppression of the cytokine cascade was given by decreased serum levels of TNF and IL-6 in CNI-1493 treated animals (TNF reduced 60% as compared to controls; IL-6 reduced 90% compared to controls; P < 0.05), and decreased levels of the acute-phase protein serum amyloid A response measured 24 h after CLP. Serum nitrites/nitrates, which give an index of NO production, were also significantly reduced (50%). Protection against CLP induced lung damage was observed as attenuation of edema and alveolar neutrophil infiltration, suppression of pulmonary TNF levels, and reduction of TUNEL-positive staining in lung. We conclude that CNI-1493 effectively inhibits the synthesis of multiple pro-inflammatory mediators and protects against death during polymicrobial sepsis.
Effects of iron on bacterial endotoxin
Authors: Roth R.I. , Panter S.S. , Zegna A.I. , Arellano F.A. , Levin J.
Published: January 2008

Bacterial endotoxin, a major modulator of morbidity and mortality during bacterial sepsis, has affinity for cations. Binding of certain cations to LPS can cause alterations of LPS size and ultrastructure, and can affect LPS biological potency. Although the addition of iron to LPS has been shown previously to result in decreased LPSmediated lethality in mice, it is not certain whether this represents a direct effect of iron on LPS biological activity. To examine this possibility, we measured binding of ferrous or ferric iron to Escherichia coli LPS in vitro. 1.5-2 moles of iron (regardless of oxidation state) was shown to bind per mole LPS. Binding of iron to LPS produced a dose-dependent decrease in two measures of LPS biological activity, activation of Limulus amebocyte lysate (which decreased to 10% of control when LPS was saturated with Fe) and stimulation of endothelial cell procoagulant activity (which decreased to 1-10% of control when LPS was saturated with Fe).
Measurement of different types of endotoxin in murine plasma by pretreatment with alkaline reagent and kinetic chromogenic Limulus test
Authors: Morita H. , Hasunuma R. , Fujihara M. , Tanaka S. , Galanos C. , Freudenberg M.A. , Morrison D.C. , Kumazawa Y.
Published: January 2008

Using a combined pretreatment methodology employing an alkaline reagent and a kinetic chromogenic Limulus test using Endospecy®, different types of endotoxins in murine plasma were able to be measured quantitatively. The detection of smooth (S)-form Salmonella abortus equi LPS (S-LPS) was approximately twice as sensitive as Helicobacter pylori LPS in aqueous solution. S-LPS added to whole mouse blood was recovered quantitatively from plasma, but the rough (R)-form S. minnesota LPS was not. The rates of clearance of S-LPS from the circulation following intravenous injection were shown to be different among BALB/c, C3H/HeN and B10Sn mice. Infection of mice with Salmonella typhimurium 3 days earlier resulted in accelerated clearance of subsequently injected LPS from the circulation. Significantly higher amounts of free non-microbe-associated LPS were detected in the blood of mice 1 h after infection with Escherichia coli O111:B4 followed by immediate treatment with ceftazidime, in comparison with non-antibiotic-treated but infected mice.
Difference in clearance of exogenously administered smooth-form LPS following host responses among normal, sensitized and LPS-tolerant mice
Authors: Morita H. , Hasunuma R. , Hoshino M. , Fujihara M. , Tanaka S. , Yamamoto S. , Kumazawa Y.
Published: January 2008

Clearance of exogenously administered Salmonella abortus equi LPS from the circulation following induction of host responses, e.g. release of soluble CD14 (sCD14) and TNFα production, were investigated. The endotoxin unit of administered LPS in plasma was monitored by a combined method. After 1 h, more than 80% of injected LPS disappeared from the circulation of normal mice at all doses except a 100 μg dose, but sCD14 in plasma could not yet be detected by Western blotting. Release of sCD14 reached a peak 9 h after LPS injection. According to pretreatment with either Propionibacterium acnes or silver nitrate, the clearance rates of exogenously added LPS from the circulation were accelerated in comparison with the rate in normal mice, but plasma TNF levels were the opposite. In LPS-tolerant mice, LPS clearance and production of TNF and sCD14 was reduced. Pretreatment with anti-CD14 mAb reduced LPS-induced TNF production but did not influence the clearance rates. Taken together, in vivo, sCD14 may not play a critical role for early LPS clearance.
Detection of lipopolysaccharides electroblotted onto membranes
Authors: Kao G.L. , Tsai C.-M.
Published: January 2008

Specific detection of LPS on nylon, as described in a previously published method (Chen W.H., Balakonis P., Tsai C-M. Detection of lipopolysaccharides blotted on nylon membranes. J Endotoxin Res 1995; 2: 405-410), has been performed with Amersham's Hybond-N. The current reformulated Hybond-N nylon membrane of Amersham did not work with the method. Magna nylon (Micron Separations Inc.) has been found as a successful substitute with results similar to that of the old Hybond-N. Incorporation of 0.05% SDS in the 25 mM Tris-192 mM glycine transfer buffer resulted in specific detection for LPS and proteins with a high content of carbohydrate. Such a phenomenon seems unique to nylon but not for PVDF and nitrocellulose.
Determinants of HRM Policies and Practices in India: An Empirical Study
Authors: Budhwar Pawan S.
Published: January 2008

Research suggests that a number of facfors influence human resource management (HRM), policies and practices. This article reports the empirical results of a study designed to examine and highlight the main contingent variables and national factors influencing Indian HRM. The investigation is based on a questionnaire survey carried out in 137 Indian firms in the manufacturing sector. The results suggest a number of significant correlations between a set of contingent variables (i.e., age, size, ownership, life cycle stage and HRM strategies of an organization, type of industry and union membership) and four HRM functions of recruitment and selection, training and development, compensation and employee communication. Similarly, four national factors (namely national culture, institutions, dynamic business environment and business sector) are suggested, which influence Indian HRM policies and practices. The study has a number of implications both for academics and practitioners. It has also opened avenues for future research.
Economic Policy in a Democratic Polity: Reforms-from Rajiv Gandhi to the Twenty-first Century
Authors: Aiyar Mani Shankar
Published: January 2008

This paper seeks to assess the economic reforms in terms of the goals and methodology of reforms. It is argued that 10 years of reforms-driven economic mallagement has failed India economically, politically and socially. Analyzing the gap between promise and performance in different sectors, the paper reflects on the performance of synthetics fibre industry in the manufacturing sector. Political empowerment of the dominant element in fibre manufacturing and spinning has matched political disempowerment of the most affected sections of the population in the decentralized cotton growing, spinning, and cloth weaving sector. If the synthetics industry is a pointer to the future of reforms, its implications for the health of our democratic polity needs to be fully understood. It is argued that economic reforms should not remain divorced from the concerns of the people. Panchayati Raj should become the center-piece of the reform process in order to ensure an economic policy that is responsive to people's needs.