bluuebunny/crossref_metadata_2025 huggingface dataset:
Dance Classes-Dancing between Classifications
Authors: Pini Maria
Published: June 2007

Women's involvement within both performance and social dance has long been trivialized by cultural analysts and social historians alike. Dance has tended then, to be dismissed as `meaningless'. However, as Angela McRobbie's work has shown, dance can take on particular significance within the context of working-class femininity. This trivialization, or regulated ignoring of social dance, can then be located as part of the wider tendency which Carolyn Steedman describes as `the tradition of cultural criticism in this country which has celebrated a kind of psychological simplicity' within working-class life (1986: 12). The treatment of the life-narrative drawn upon in this article seeks to illustrate the complex meanings an involvement in dance can have within the context of a working-class girlhood and womanhood, and thus aims at highlighting how involvement within a popular cultural practice such as dance relates to the constitution of subjectivity.
Bleeding out Loud: Communication about Menstruation
Authors: Kissling Elizabeth Arveda
Published: June 2007

Girls in the USA receive mixed messages about menarche: menarche is traumatic and upsetting-but act normal; menarche is an overt symbol of sexual maturity-but also a mysterious, secret event. Girls I interviewed reported another-dualism in their menstrual education. They perceive a clear distinction between two kinds of menstrual knowledge: scientific knowledge about the anatomy and physiological functioning, and what they termed `realistic', pragmatic knowledge about managing the lived experience of menstruation. Using methods of critical, feminist analysis, I examine social texts of menstrual socialization, including girls' conversation about their menstrual education; their mothers' discussions of the preparation for menstruation that girls received; and instructional materials presented in their health education classes, to discuss the communication and the silences of contemporary menstrual socialization, and to suggest possibilities for transformative language and action to teach girls about menstruation in ways that can help mend dualisms.
Never Innocent: Breasted Experiences in Women's Bodily Narratives of Puberty
Authors: Lee Janet
Published: June 2007

In this article I focus on the social construction of the female breast as a central aspect of body politics, emphasizing how the breasted female body becomes invested with certain properties and is inserted into regimes of truth, knowledge and power. I share data from an oral history project of women's experience of menarche or first menstrual period and explore how cultural scripts associated with developing female breasts are negotiated and lived by women through processes of compliance and resistance. I suggest that in western societies, the ambivalence women remember feeling about their developing breasts is a result of the simultaneous experience of breasts as signifying both disempowerment and sexual authority.
Multiple Chemical Sensitivity, Culture and Delegitimization: A Feminist Analysis
Authors: Gibson Pamela Reed
Published: June 2007

The condition referred to as multiple chemical sensitivity (MCS), chemical injury (CI), environmental illness (El), 20th-century disease or toxic encephalopathy has generated controversy because victims' claims of harm from low levels of chemicals in the environment conflict with general mainstream belief in the safety of these substances. Mainstream (western) culture's response to persons with MCSICI has interacted with sexism, served to delegitimize victims thereby reducing their power, reduced the victim's context (because of forced isolation), and jeopardized jobs and relationships. This article will analyze the situation of the chemically injured person and culture's response from a feminist perspective, and discuss cultural delegitimization as a process used by a chemical culture to neutralize the message of the person with MCSICI, i.e., that the environment is unsafe for her/him, and, by extension, for others.
`It's a Nice Day for a White Wedding': The Debutante Ball and Constructions of Femininity
Authors: Harrison Lyn
Published: June 2007

This article focuses on three young women's experiences in relation to their participation in a debutante ball in their penultimate year of secondary school. The debutante ball is used to explore the place of desire and fantasy in the lives of young women and the ways in which their bodies are disciplined, within its rituals, into the normalizing discourses and discursive practices of femininity and heterosexuality. The analysis makes evident the discursive construction of a femininity/feminism opposition and the effect this opposition has on these young women's perceptions of the place of feminism in their lives. Practices of femininity are related to pleasure whereas feminism is often perceived in terms of a coercive form of puritanism.
Invited review: Enteric bacteria, lipopolysaccharides and related cytokines in inflammatory bowel disease: biological and clinical significance
Authors: Caradonna L. , Amati L. , Magrone T. , Pellegrino N.M. , Jirillo E. , Caccavo D.
Published: June 2007

Ulcerative colitis (UC) and Crohn's disease (CD) [inflammatory bowel disease (IBD)] are both characterized by an exaggerated immune response at the gut associated lymphoreticular tissue level. Such an abnormal and dysregulated immune response may be directed against luminal and/or enteric bacterial antigens, as also supported by murine models of inflammatory bowel disease (IBD) caused by organisms such as Citrobacter rodentium and Helicobacter hepaticus.

Bacterial endotoxins or lipopolysaccharides (LPS) have been detected in the plasma of IBD patients and an abnormal microflora and/or an increased permeability of the intestinal mucosa have been invoked as cofactors responsible for endotoxemia. At the same time, the evidence that phagocytosis and killing exerted by polymorphonuclear cells and monocytes and the T-cell dependent antibacterial activity are decreased in IBD patients may also explain the origin of LPS in these diseases.

In IBD, pro-inflammatory cytokines and chemokines have been detected in elevated amounts in mucosal tissue and/or in peripheral blood, thus suggesting a monocyte/macrophage stimulation by enteric bacteria and/or their constituents ( e.g. LPS).

On these grounds, in experimental models and in human IBD, anti-cytokine monoclonal antibodies and interleukin receptor antagonists are under investigation for their capacity to neutralize the noxious effects of immune mediators. Finally, the administration of lactobacilli is beneficial in human IBD and, in murine colitis, this treatment leads to a normalization of intestinal flora, reducing the number of colonic mucosal adherent and translocated bacteria.
Bacterial endotoxin (lipopolysaccharide) stimulates the rate of iron oxidation
Authors: Roth Robert I. , Panter Scott S. , Zegna Angelo I. , Levin Jack
Published: June 2007

Bacterial endotoxin (lipopolysaccharide) has affinity for a number of cations, including iron. Previous investigations have demonstrated that lipopolysaccharide can affect the oxidation rate of iron; heme-bound ferrous iron in hemoglobin is oxidized to ferric iron when hemoglobin binds lipopolysaccharide. In the present study, we directly examined the interaction between lipopolysaccharide and iron. Lipopolysaccharide caused a concentration-dependent increase in the rate of iron oxidation, with up to a 23-fold increase in oxidation in the presence of 200 µg/ml Escherichia coli lipopolysaccharide. This effect was seen both with several carbohydrate-rich smooth lipopolysaccharides and also with carbohydrate-poor rough lipopolysaccharide. Extensively deacylated rough lipopolysaccharide had no effect, suggesting a role of the fatty acid components of lipopolysaccharide in this process. Purified lipid A produced inconsistent results: some preparations stimulated iron oxidation and others did not. A series of sugars, starches and a preparation of purified O-chain polysaccharide (the carbohydrate portion of the lipopolysaccharide macromolecule) had no effect on the rate of iron oxidation, whereas phospholipid-enriched brain tissue extracts (similar to the lipid A component of lipopolysaccharide) stimulated oxidation. We conclude that the lipid moiety of bacterial lipopolysaccharide is responsible for the stimulation of iron oxidation. This process may contribute to the ability of lipopolysaccharide to cause oxidation of heme-bound iron in hemoglobin.
Structural requirements of muramylpeptides for induction of Toll-like receptor 2-mediated NF-κB activation in CHO cells
Authors: Yoshimura Atsutoshi , Takada Haruhiko , Kaneko Takashi , Kato Ihachi , Golenbock Douglas , Hara Yoshitaka
Published: June 2007

We previously demonstrated that Gram-positive bacteria activated immune cells via CD14 and Toll-like receptor 2 (TLR2). Although peptidoglycan, a major constituent of the bacterial cell wall, substituted for whole organisms, the essential structure of muramylpeptides required to stimulate the cells is not clear. We further investigated the critical determinant for recognition by CD14 and TLR2. Chinese hamster ovary (CHO) fibroblasts, which do not express a functional TLR2 transcript, were transfected with TLR2 or TLR4. These cells were exposed to freeze-dried Staphylococcus epidermidis and were subsequently subjected to the pro-inflammatory transcription factor nuclear factor-κB (NF-κB)-dependent CD25 expression assay. Heterologous expression of human TLR2, but not TLR4, in CHO cells conferred immune responsiveness to freeze-dried S. epidermidis. A preparation of peptidoglycan from S. epidermidis substituted for whole organisms. Staphylococcus aureus lytic enzyme-digested product (SEPS) from peptidoglycan retained the activity, but hydrolysis of the glycan backbone in SEPS by M-1 endo- N-acetylmuramidase resulted in loss of the activity. These findings showed that cellular activation by Gram-positive cell wall components was mediated by TLR2, but not TLR4, and indicated that the glycan backbone of peptidoglycan is critical for TLR2-mediated NF-κB activation.
Differential roles of TLR2 and TLR4 in the host response to Gram-negative bacteria: lessons from a lipopolysaccharide-deficient mutant of Neisseria meningitidis
Authors: Ingalls Robin R. , Lien Egil , Golenbock Douglas T.
Published: June 2007

The inflammatory response to bacterial infections plays an important role in the detection and elimination of invading micro-organisms. Various components of the bacterial cell wall are capable of activating this pro-inflammatory response. In the case of Gram-negative bacteria, lipopolysaccharide (LPS) is the dominant trigger, although other bacterial factors are also capable of activating this systemic inflammatory response. Recently, Toll-like receptors (TLRs) have been implicated in host responses to bacterial pathogens. Specifically, TLR4 mediates LPS responses while TLR2 plays a broader role in the recognition of a variety of bacteria and bacterial antigens. The experiments in this study were designed to examine the role of Gram-negative cell wall components, other than LPS, and their cellular receptors in the host response to infection using an LPS-deficient mutant of Neisseria meningitidis. Although less potent than the parental strain, we found the LPS-deficient mutant to be a capable inducer of the inflammatory response in a variety of cell types. Moreover, cellular activation by this mutant required expression of CD14 and TLR2.
A horseshoe crab receptor structurally related to Drosophila Toll
Authors: Inamori Kei-ichiro , Koori Kumiko , Mishima Chiemi , Muta Tatsushi , Kawabata Shun-ichiro
Published: June 2007

Innate immunity against microbial pathogens relies on the pattern recognition of cell wall components on invading microbes. Recent evidence has shown that a mammalian Toll-like receptor (TLR) is activated by bacterial lipopolysaccharides (LPS). The innate immunity in invertebrates is also triggered by LPS, as seen in the hemolymph coagulation in horseshoe crab. We report the cloning of a TLR from the Japanese horseshoe crab Tachypleus tridentatus. A cDNA coding for Tachypleus Toll was isolated from a hemocyte cDNA library and the open reading frame codes for a proprotein including a signal sequence. Like Drosophila Toll, Tachypleus Toll is a type I transmembrane protein with an extracellular domain consisting of two leucine-rich repeats flanked by two cystein-rich clusters and a cytoplasmic domain exhibiting striking similarity with the cytoplasmic domain of interleukin-1 receptor. Tachypleus Toll is most similar to Drosophila Toll in the domain architecture and the overall length.
Interferon-γ counteracts reduced endotoxin responsiveness of whole blood following trauma and cardiopulmonary bypass
Authors: Flohé Sascha , Börgermann Jochen , Lim Lucy , Schade Fritz-Ulrich
Published: June 2007

Accidental as well as surgical trauma has been reported to cause reduced endotoxin responsiveness of blood in terms of cytokine production. In this study, the effect of interferon-γ (IFN-γ) on tumour necrosis factor-α (TNF-α)-producing capacity of whole blood after severe trauma and cardiac surgery was investigated. Blood samples of severely injured patients were collected at the first day after trauma and of cardiac surgery patients before, 4 h and 2 days after cardiopulmonary bypass (CPB). The blood samples were incubated with INF-γ (0—100 U/ml) for 20 h and subsequently lipopolysaccharide (LPS)-induced TNF-α production was determined. Compared to healthy donors, LPS-induced TNF-α production was significantly reduced in blood cultures of trauma patients on day 1 after trauma and 4 h after CPB. Pre-incubation with IFN-γ in vitro increased endotoxin-induced TNF-α production in volunteers' and all patients' blood specimens in a dose-dependent manner. IFN-γ prompted an elevation of cytokine synthesis in CPB patients' blood which equalled that of volunteers, whereas it caused a lower rise in TNF-α production in blood of multiply injured patients, reaching levels of untreated donors only after incubation with 100 U/ml IFN-γ. These experiments show that hyporesponsiveness of whole blood induced by trauma or cardiac surgery with CPB is not irreversible, but can be counteracted by the immunostimulant IFN-γ. IFN-γ, therefore, could be applied clinically in trauma patients or after cardiac surgery to prevent or to resolve infection complications.
Chemical structure and biological activity of a lipid A component from Helicobacter pylori strain 206
Authors: Suda Yasuo , Kim Yong-Min , Ogawa Tomohiko , Yasui Norio , Hasegawa Yushi , Kashihara Wataru , Shimoyama Takashi , Aoyama Kazue , Nagata Kumiko , Tamura Toshihide , Kusumoto Shoichi
Published: June 2007

The chemical structure of a lipid A, which was obtained as a minor component from lipopolysaccharide of Helicobacter pylori strain 206-1, was determined to be a glucosamine β-(1-6) disaccharide 1-(2-aminoethyl)phosphate acylated by ( R)-3-hydroxyoctadecanoic acid, ( R)-3-hydroxyhexadecanoic acid, and ( R)-3-(octadecanoyloxy)octadecanoic acid at the 2-, 3- and 2′positions, respectively. Compared with the other major lipid A from the same strain, which was previously reported [Suda Y, Ogawa T, Kashihara W et al. Chemical structure of lipid A from Helicobacter pylori strain 206-1 lipopolysaccharide. J Biochem 1997; 121: 1129—1133], the structure was very similar with one exception. An ( R)-3-hydroxyhexadecanoic acid was present at the 3-position of the novel lipid A component. The structure is apparently identical to one of the proposals by Moran et al. [Moran AP, Lindner B, Walsh EJ. Structural characterization of the lipid A component of Helicobacter pylori rough- and smooth-form lipopolysaccharides. J Bacteriol 1997; 179: 6453—6463], who concluded the same structure as the so-called major lipid A from the H. pylori strain NCTC 11637 but without isolating a homogenous component. The endotoxic properties and pro-inflammatory cytokine-inducing activities of this novel tetra-acyl type lipid A were lower than those of previously reported major tri-acyl type lipid A.
Synthesis and immunochemical characterization of neoglycoproteins containing epitopes of the inner core region of Pseudomonas aeruginosa RNA group I lipopolysaccharide
Authors: Reiter Andreas , Brade Lore , Sanchez-Carballo Patricia , Brade Helmut , Kosma Paul
Published: June 2007

The disaccharide allyl 2-acetamido-2-deoxy-α-D-galactopyranosyl-(1→3)-7- O-carbamoyl-L- glyceroα-D- manno-heptopyranoside 5 (GalNAc-cmHep-allyl) was synthesized starting from 1 and 2. Compound 5, cmHep-allyl and the disaccharide cmHep-(1→3)-Hep-allyl were converted into cysteamine-spacered derivatives and conjugated to bovine serum albumin (BSA) to yield the neoglycoconjugates 7—9, respectively. These conjugates were used to immunize mice and to prepare monoclonal antibodies (mAbs) which were characterized in comparison to mAbs obtained after immunization with heat-killed Pseudomonas aeruginosa strain H4. Two antibodies obtained after immunization with the neoglycoconjugates bound strongly to cmHep-BSA and with lower affinity to cmHep-Hep-BSA but did not bind to GalNAc-cmHep-BSA or to H4 LPS. Another antibody obtained after immunization with heat-killed bacteria bound to LPS and GalNAc-cmHep-BSA but not to cmHep-Hep-BSA or cmHep-BSA
Up-regulation of gastric mucosal inflammatory responses to Helicobacter pylori lipopolysaccharide by aspirin but not indomethacin
Authors: Slomiany B.L. , Piotrowski J. , Slomiany A.
Published: June 2007

Infection with Helicobacter pylori and the use of non-steroidal anti-inflammatory drugs (NSAIDs) are recognized as the two primary factors in the etiology of gastric disease. In this study, we applied the animal model of H. pylori-induced gastritis to assess the influence of NSAIDs on the course of mucosal inflammatory responses to H. pylori. Two days following intragastric application of H. pylori lipopolysaccharide, rats were divided into groups and administered daily for up to 8 days either indomethacin, aspirin or the vehicle, and their gastric mucosal tissue subjected to histological and biochemical assessment. H. pylori lipopolysaccharide elicited within 2 days a pattern of acute mucosal inflammatory responses accompanied by a massive epithelial cell apoptosis, and a marked increase in the expression of membrane-bound and soluble forms of TNF-α. The extent of mucosal inflammatory involvement reached a maximum by the 4th day and showed a decline by the 10th day; this was reflected in a 38.1% reduction in apoptosis, a 53.2% decline in membrane-bound TNF-α and a 63.8% decrease in soluble TNF-α. Compared to the vehicle controls, aspirin caused a 36.2% increase in the severity of the mucosal inflammatory involvement by the second day of administration and a 25.9% increase in the inflammatory involvement occurred by the 8th day; this effect of aspirin was accompanied by a significant (54.5%) induction in apoptosis, a 58.2% decline in membrane-bound TNF-α and a 61% increase in soluble TNF-α. In contrast, administration of indomethacin evoked only a marginal increase (5—7%) in apoptosis, and caused no discernible changes in the severity of gastric mucosal involvement and the expression of TNF-α forms elicited by H. pylori lipopolysaccharide. The findings indicate that aspirin, but not indomethacin, increases the severity of gastric mucosal inflammatory responses to H. pylori. This detrimental influence of aspirin appears to result from up-regulation in the mucosal expression of soluble form of TNF-α, which leads to the amplification of apoptotic events that potentiate gastric mucosal inflammatory reaction to H. pylori.
The expression of membrane-bound CD14 renders mouse B-1 cells susceptible to LPS
Authors: Sugiyama Tsuyoshi , Koide Naoki , Chakravortty Dipshika , Kato Yutaka , Mu Mya Mya , Yoshida Tomoaki , Yokochi Takashi
Published: June 2007

The surface expression of CD14 on mouse B-1 cells and its role on their response to lipopolysaccharide (LPS) were studied by using the murine TH2.52 B-1 cell line and peritoneal B-1 cells. TH2.52 cells with the B-1 phenotype were found to express membrane-bound CD14. Furthermore, CD14 was expressed on physiological peritoneal CD5 + B-1 cells. The stimulation of CD14-expressing TH2.52 cells with a low concentration of LPS resulted in the activation of nuclear factor (NF)-κB and a mitogen-activated protein kinase (MAPK). The LPS-induced NF-κB and MAPK activation was markedly inhibited by anti-CD14 antibody. These results suggest that B-1 cells may respond to LPS via membrane-bound CD14.
Decay-accelerating factor (DAF/CD55) is a functional active element of the LPS receptor complex
Authors: Heine H. , Ulmer A.J. , El-Samalouti V.T. , Lentschat A. , Hamann L.
Published: June 2007

Previously, we identified an 80 kDa membrane protein (LMP80) that is capable of binding to LPS and lipid A in the presence of LBP and sCD14. LMP80 could also be detected after immuno-coprecipitation of cell membranes with LPS and lipid A, indicating a physical contact of LMP80 and LPS/lipid A. Further analysis and peptide sequencing revealed that LMP80 is identical to CD55 (decay accelerating factor, DAF), a regulatory molecule of the complement cascade. Transfection of LPS-hyporesponsive Chinese hamster ovary (CHO) cells with human CD55 resulted in the translocation of NF-κB upon stimulation with LPS or lipid A. Our results demonstrate a new functional role of CD55 as a molecule able to mediate LPS-induced activation of cells that may be part of a multimeric LPS receptor complex.
Involvement of TLR4/MD-2 complex in species-specific lipopolysaccharide-mimetic signal transduction by Taxol
Authors: Kawasaki Kiyoshi , Akashi Sachiko , Shimazu Rintaro , Yoshida Takashi , Miyake Kensuke , Nishijima Masahiro
Published: June 2007

Taxol, an antitumor agent derived from a plant, mimics the action of lipopolysaccharide (LPS) in mice, but not in humans. The LPS-mimetic activity of Taxol is not observed in LPS-hyporesponsive C3H/HeJ mice which possess a point mutation in Toll-like receptor 4 (TLR4); therefore, TLR4 appears to be involved in both Taxol and LPS signaling. In addition, TLR4 was recently shown to physically associate with MD-2, a molecule that confers LPS-responsiveness on TLR4. Here we examined whether or not TLR4/MD-2 complex mediates a Taxol-induced signal by using transformants of the mouse pro-B cell line, Ba/F3, expressing mouse TLR4 alone, both mouse TLR4 and mouse MD-2, and both mouse MD-2 and mouse TLR4 lacking the cytoplasmic portion. Our results demonstrated that co-expression of mouse TLR4 and mouse MD-2 was required for Taxol responsiveness, and that the TLR4/MD-2 complex is the shared molecule in Taxol and LPS signal transduction in mice. We also found that mouse MD-2, but not human MD-2, is involved in Taxol signaling, suggesting that MD-2 is responsible for the species-specific responsiveness to Taxol.
Characterization of the physiological substrate for lipopolysaccharide heptosyltransferases I and II
Authors: Gronow Sabine , Oertelt Clemens , Ervelä Elise , Zamyatina Alla , Kosma Paul , Skurnik Mikael , Holst Otto
Published: June 2007

L- Glycero-D- manno-heptopyranose is a characteristic compound of many lipopolysaccharide (LPS) core structures of Gram-negative bacteria. In Escherichia coli two heptosyltransferases, namely WaaC and WaaF, are known to transfer L- glycero-D- manno-heptopyranose to Re-LPS and Rd 2-LPS, respectively. It had been proposed that both reactions involve ADPL- glycero-D- manno-heptose as a sugar donor; however, the structure of this nucleotide sugar had never been completely elucidated. In the present study, ADPL- glycero-D- manno-heptose was isolated from a heptosyltransferase-deficient E. coli mutant, and its structure was determined by nuclear magnetic resonance spectroscopy and matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry as ADPL- glycero-β-D- manno-heptopyranose. This compound represented the sole constituent of the bacterial extract that was accepted as a sugar donor by heptosyltransferases I and II in vitro .
The hepatocyte as a microbial product-responsive cell
Authors: Vodovotz Yoram , Shubing Liu  , McCloskey Carol , Shapiro Richard , Green Angela , Billiar Timothy R.
Published: June 2007

Much research has focused on the responses to microbial products of immune cells such as monocytes, macrophages, and neutrophils. Although the liver is a primary response organ in various infections, relatively little is known about the antimicrobial responses of its major cell type, the hepatocyte. It is now known that the recognition of bacteria occurs via cell-surface proteins that are members of the Toll-like receptor (TLR) family. In addition, lipopolysaccharide (LPS) is bound by circulating LPS-binding protein (LBP) and presented to cell-surface CD14, which in turn interacts with TLR and transduces an intracellular signal. We investigated the CD14 and TLR2 responses of whole liver and isolated hepatocytes, and demonstrated that these cells can be induced to express the molecules necessary for responses to both Gram-positive and Gram-negative bacteria. Our findings may have clinical implications for pathological states such as sepsis.
The role of TLR4 in endotoxin responsiveness in humans
Authors: Schwartz David A.
Published: June 2007

Despite the tremendous inter-individual variability in the response to inhaled toxins, we simply do not understand why certain people develop disease when challenged with environmental agents and others remain healthy. To address this concern, we investigated whether the Toll-4 (TLR4) gene, that has been shown to affect lipopolysaccharide (LPS) responsiveness in mice, underlies the variability in airway responsiveness to inhaled LPS in humans. Here we show that common, co-segregating missense mutations (Asp299Gly and Thr399Ile) in the extracellular domain of the TLR4 receptor are associated with a significantly blunted response to inhaled LPS in 83 humans. Transfection of THP-1 cells demonstrates that the Asp299Gly mutation (but not the Thr399Ile mutation) interrupts TLR4-mediated LPS signaling. Moreover, the wild-type allele of TLR4 rescues the LPS hyporesponsive phenotype in either primary airway epithelial cells or alveolar macrophages obtained from individuals with the TLR4 mutations. Our findings provide the first genetic evidence that common mutations in TLR4 are associated with differences in LPS responsiveness in humans, and demonstrate that gene sequence changes can alter the ability of the host to respond to environmental stress.
Complement-dependent platelet degradation and anaphylactoid shock in mice induced by lipopolysaccharide carrying the mannose homopolymer
Authors: Endo Yasuo , Yokochi Takashi , Matsushita Misao , Fujita Teizo , Takada Haruhiko
Published: June 2007

Intravenous injection of specified bacterial lipopolysaccharides (LPSs) induced anaphylactoid shock in mice of various strains, including LPS-resistant C3H/HeJ. The reaction was accompanied by occasional mortality of mice within 1 h. Prior to shock, rapid accumulation of blood platelets in the lungs and liver followed by degradation of platelets (or release of their contents) and tissue destruction were observed. In this study, LPS specimens carrying mannose-homopolymer (MHP), which markedly activate the human complement system through the lectin pathway, induced marked platelet degradation and anaphylactoid shock in BALB/c mice. In contrast, in C5-deficient DBA/2 mice, the platelet degradation and anaphylactoid reactions did not occur. Anti-complement agent K-76 COOH (C5 inhibitor) protected BALB/c mice from mortality in the anaphylactoid reaction. K-76 COOH also inhibited platelet degradation, but not accumulation, induced by LPS in mice. Based on these findings, we postulated that strong complement activation by specified LPS preparations induced degradation of platelets that have accumulated in the lungs and liver, resulting in acute inflammation accompanied by severe tissue destruction, especially in the lungs, which in turn leads to anaphylactoid reaction.
Macrophage migration inhibitory factor (MIF) modulates innate immune responses induced by endotoxin and Gram-negative bacteria
Authors: Roger Thierry , Glauser Michel P. , Calandra Thierry
Published: June 2007

Discovered in the early 1960s as a T-cell cytokine, MIF has emerged to be an important mediator of the innate immune system. MIF was identified recently to be released by a vast array of cells, including monocytes/macrophages, T-cells, B-cells, endocrine cells and epithelial cells in response to infection and stress. Bacteria, microbial toxins and cytokines have been shown to be powerful inducers of MIF secretion by macrophages. MIF stimulates the expression of pro-inflammatory mediators by immune cells and functions to counterbalance the anti-inflammatory and immunosuppressive effects of glucocorticoids. Like TNF and IL-1, MIF plays an important role in host responses to infection. Recombinant MIF was found to exacerbate lethal endotoxemia or bacterial sepsis when co-injected with LPS or Escherichia coli in mice. Conversely, MIF knockout mice or mice treated with anti-MIF antibodies were protected from shock induced by LPS, staphylococcal exotoxins or bacterial peritonitis, even when anti-MIF therapy was started after the onset of infection. Given the central role played by MIF in innate immune responses against microbial pathogens and in the regulation of inflammatory responses, pharmacological modulation of MIF production or neutralization of MIF activity could have broad clinical applications and may offer new treatment options for the management of patients with severe sepsis or septic shock.
Modulation of endotoxin-induced cardiopulmonary dysfunction by S-nitroso-albumin
Authors: Gluckman Tracy L. , Grossman Jeffrey E. , Folts John D. , Kruse-Elliott Kris T.
Published: June 2007

Nitric oxide (NO) is an endogenous vasodilator and modulator of inflammation. During endotoxemia, the beneficial effects of NO are overwhelmed by the inflammatory cascade, resulting in a functional depletion of NO. S-nitroso-albumin ( S-NO-alb) exists as a novel and highly stable NO thiol complex that slowly releases NO into the vascular micro-environment. Using a porcine model, we examined the ability of intravenous S-NO-alb to modulate cardiopulmonary dysfunction characteristic of endotoxemia. Pigs were anesthetized, instrumented for standard cardiopulmonary function measurements, and randomly assigned to receive: (i) albumin + saline; (ii) albumin + LPS; or (iii) S-NO-alb + LPS. Cardiopulmonary parameters were evaluated every 30 min and ex vivo phorbol myristate acetate (PMA)-stimulated superoxide release was serially determined as a marker of in vivo neutrophil priming. Lung myeloperoxidase (MPO) activity was measured as a marker of neutrophil migration into the lung. LPS-induced cardiopulmonary dysfunction was characterized by a sustained elevation in mean pulmonary arterial pressure, pulmonary vascular resistance, and peak intratracheal pressure, as well as a reduction in cardiac index, stroke volume index and PaO2over 6 h. Pretreatment with S-NO-alb attenuated LPS-induced cardiopulmonary dysfunction without adversely affecting systemic hemodynamics. Moreover, S-NO-alb blunted the LPS-induced hypoxemic response and reduced neutrophil activation. S-NO-alb did not, however, attenuate LPS-induced increases in lung MPO. Our results suggest that S-NO-alb can selectively modulate endotoxin-induced pulmonary dysfunction, attenuate neutrophil priming and block the early mortality (40%) in this model.
The biological activity of a liposomal complete core lipopolysaccharide vaccine
Authors: Erridge Clett , Stewart John , Bennett-Guerrero Elliott , McIntosh Thomas J. , Poxton Ian R.
Published: June 2007

A vaccine that induces humoral immunity to lipopolysaccharide (LPS), while remaining non-pyrogenic should be beneficial, as high levels of antibodies against LPS are associated with a reduced risk of adverse outcome. However, pure LPS or bacteria expressing LPS are generally considered too toxic to be used as vaccines. Recently, a novel, immunogenic complete core lipopolysaccharide vaccine has been described, which has been designed to prevent endotoxin-related inflammatory reactions in surgical and high-risk hospitalized patients. In vivo studies have shown that while administration of the vaccine to rabbits results in no toxicity over 7 days, it does induce significantly enhanced antibody responses towards a broad range of clinically relevant Gram-negative LPSs. Here we show that encapsulation of the four complete core LPS types Escherichia coli K12, Escherichia coli R1, Bacteroides fragilis and Pseudomonas aeruginosa into liposomes greatly reduces the ability of a given amount of LPS to induce TNF-α production in vitro from human monocytes. In contrast to previous studies of liposomal LPS, we demonstrate a reduction in activity of approximately 100,000-fold; a reduction approximately 100—1,000-fold more than that previously described. The signalling by the liposomal LPS appears to be entirely dependent on serum factors, though this can be partially restored by soluble CD14 or, to a lesser extent, by lipopolysaccharide binding protein. Time-course experiments reveal that liposomal LPS signalling shows similar kinetics to pure LPS signalling. Therefore, as well as inducing specific antibody responses, liposomal LPS demonstrates characteristics suitable for use as a vaccine to be used in human beings.
Staphylococcal enterotoxin B induces hepatic injury and lethal shock in endotoxin-resistant C3H/HeJ mice despite a deficient macrophage response
Authors: Yasuda Shigeo , Nagaki Masahito , Moriwaki Hisataka
Published: June 2007

Bacterial toxins, including endotoxin/LPS as well as superantigens, are major causative agents of multi-organ failure associated with sepsis and liver disease. However, the precise mechanisms initiating cell activation by the toxins have not been clarified. We compared lethal shock and cytokine production in response to LPS with responses to the superantigen staphylococcal enterotoxin B (SEB) in both LPS-responsive C3H/HeN mice and LPS-hyporesponsive C3H/HeJ mice treated with D-galactosamine (GalN). LPS was not lethal and did not induce production of TNF-α in C3H/HeJ mice. In contrast, SEB produced lethal shock associated with liver failure and induced cytokines such as TNF-α, IFN-γ, and IL-2 in both C3H/HeN and C3H/HeJ mice. Peritoneal macrophages from C3H/HeJ mice did not produce TNF-α in vitro in response to SEB or LPS. However, no significant difference was observed in production of TNF-α in response to stimulation in vitro by SEB between C3H/HeN and C3H/HeJ splenic lymphocytes. We have demonstrated that SEB causes lethal toxicity associated with liver injury in LPS-hyporesponsive C3H/HeJ mice and that as the underlying mechanism, the normal T-cell function in these mice still maintained the sensitivity to SEB since the genetic defect of C3H/HeJ mice unresponsive to LPS and SEB is restricted in macrophages/monocytes and does not extend to T cells.
Lipopolysaccharide binds to and activates A1 adenosine receptors on human pulmonary artery endothelial cells
Authors: Neely Wilson Constance , Batra Vinod K.
Published: June 2007

Previously, it was reported that A1 adenosine receptor antagonists prevent endotoxin-inducedacute lung injury and pulmonary arterial endothelial cell damage. In competition radioligand binding experiments in membranes prepared from human pulmonary artery endothelial cells (PAECs), lipopolysaccharides (LPSs) of Escherichia coli, Salmonella typhimurium, Klebsiella pneumoniae, and Pseudomonas aeruginosa displaced the binding of a selective A adenosine receptor antagonist [125I]-BWA844U (IC 50 values: 195 ng/ml, 290 ng/ml, 602 ng/ml, and 6931 ng/ml, respectively)in a dose-dependent, competitive manner. There was no displacement of this radioligand by enterotoxin (≤ 10 μg/ml), diphosphoryl lipid A (≤ 10μg/ml), and glycolipids, monosialoganglioside(≤ 1μg/ml), lactocerebroside (≤ 100μg/ml), or NBD galactocerebroside (≤ 100 μg/ml). Based on calculated IC values, LPS ( E. coli, IC50 111 ng/ml) 50 6 displaced the selective A1 adenosine receptor agonist, [3H]-2-chloro, N -cyclopentyladenosine (CCPA) in human PAECs with a potency profile, CCPA > LPS > 2-phenylaminoadenosine (CV 1808), a selective A2 adenosine receptor agonist. The potency profile for displacement of the selective A μ 2a adenosine receptor agonist [ 3H]-CGS 21680 was CV 1808 > CCPA. LPS ( E. coli 0.1 pg/ml—10 g/ml) did not displace [3H]-CGS 21680 binding. In human PAECs, IL-6 and TXA2 release induced by LPS (0—1 μg/ml) or CCPA (0—1 μM) at high doses was significantly reduced by the selective A1 adenosine receptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine(DPCPX; 1 μM). These data suggest that LPS binds to and activates A1 adenosine receptors on human PAECs to induce the release of IL-6 and TXA 2. Activation of A1 adenosine receptors on human PAECs by LPS, may contribute to the pathophysiology of acute lung injury associated with Gram-negative septicemia and endotoxemia.
The central role of thromboxane and platelet activating factor receptors in ex vivo regulation of endotoxin-induced monocyte tissue factor activity in human whole blood
Authors: Eilertsen Karl-Erik , Østerud Bjarne
Published: June 2007

Expression of tissue factor (TF) by activated monocytes may initiate thrombotic episodes associated with diseases, such as thrombosis and atherosclerosis. In this study, steps in the regulatory pathways of lipopolysaccharide (LPS)-induced monocyte TF activity and released TNF-α in human whole blood were probed for using an array of inhibitors, comprising specific inhibitors of cytosolic phospholipase A2 (PLA2) (AACOCF3), secretory PLA (SB-203347), protein kinase (PK) (staurosporine), PKC (GF109203; BIM), and serine protease (Pefabloc SC), antagonists of thromboxane prostanoid (TP) receptor (R) (SQ-29548), platelet activating factor (PAF) R (BN-52021), leukotriene B4 R (SC-41930), serotonin R (cyproheptadine), fibronectin/fibrinogen R (RGDS), and finally, creatine phosphate/creatine phosphokinase (CP/CPK) which removes ADP. Whereas when added alone neither of these agents significantly inhibited LPS-induced TF or TNF-α, when presented as a reference cocktail comprising all the agents, TF activity and TNF-α were reduced by 77% and 49%, respectively. By subsequently testing a series of incomplete inhibitory cocktails equal to the reference except for deleted single agents or combinations of two or three active agents, the inhibitory effect of the reference cocktail could be shown to depend on the presence of the protease inhibitor and the thromboxane A2 and PAF antagonists.
Review paper: LPS in microbial pathogenesis: promise and fulfilment
Authors: Beutler Bruce
Published: June 2007

Since LPS was discovered, all inquiry i n the field was motivated by two articles of faith. First, it was held that LPS was an important factor in microbial pathogenesis. Second, it was held that the responses elicited by numerous molecules of microbial origin might be formally similar to the responses elicited by LPS. The identification of TLR4 as the core transducer of LPS responses, and the added discovery that other TLR paralogs sense other microbial products, has strongly validated these cherished beliefs. M oreover, it has amalgamated many separate lines of inquiry, and focused attention on the how the innate immune system perceives infection.
Monoclonal antibodies against 3-deoxy-α-D-manno-oct-2-ulosonic acid (Kdo) and D-glycero-α-D-talo-oct-2-ulosonic acid (Ko)
Authors: Brade Lore , Gronow Sabine , Wimmer Norbert , Kosma Paul , Brade Helmut
Published: June 2007

Monoclonal antibodies (mAbs) were obtained after immunization of mice with neoglycoconjugates containing as a ligand the disaccharide Kdo(2→ 4)Ko or Ko(2→4)Kdo, representing structural elements of the core region of lipopolysaccharides (LPSs) from Acinetobacter haemolyticus and Burkholderia cepacia, respectively. One antibody, S67-9, bound with high specificity to Ko(2→ 4)Kdo-BSA showing no cross reactivity with Kdo(2→ 4)Kdo-BSA and the other antigens tested. A second mAb, S68-12, bound preferentially to Kdo(2→ 4)Ko-BSA but cross reacted with Ko(2→ 4)Kdo-BSA and Kdo(2→ 4)Kdo-BSA. A third mAb, S67-27, was found to bind Kdo monosaccharide. Although mAbs S67-9 and S68-12 did not bind to LPS of Burkholderia or Acinetobacter as expected, the mAbs will be useful tools in studying the biosynthesis of LPS containing Ko.
Antagonism of in vivo and ex vivo response to endotoxin by E5564, a synthetic lipid A analogue
Authors: Rossignol Daniel P. , Lynn Melvyn
Published: June 2007

E5564, a synthetic lipid A analogue, is a selective, highly active antagonist of endotoxin-mediated activation of immune cells. Preclinical research has indicated that E5564 can block endotoxin-mediated induction of cytokines and endotoxin or Gram-negative bacterial-induced death in animal models. Recent phase I clinical trials have focused on the ability of E5564 to block responsiveness to endotoxin. This was done in two ways: in vivo challenge of human volunteers with 4 ng/kg endotoxin, and by use of an ex vivo assay which utilizes blood drawn from volunteers administered E5564 and challenged with endotoxin at concentrations that ranged from 50 pg/ml to 10 ng/ml. In vivo, 100 μg of E5564 completely blocked signs, symptoms and cytokines induced by concomitantly-administered endotoxin. In contrast, subjects receiving a 50 μg dose of E5564 demonstrated a graded response; cytokines were inhibited 95%, but many signs and symptoms of endotoxemia were still evident. E5564 demonstrated a long pharmacokinetic half-life (> 30 h); however, ex vivo analysis indicated that while single doses of 350 μg induced a nearly complete block of the effects of 1 ng/ml endotoxin immediately upon E5564 administration, antagonistic activity declined rapidly (t 1/2 < 1 h). Similar results were obtained in vivo using a delayed endotoxin challenge. These results have driven us to examine antagonistic activity of E5564 in vivo and ex vivo after administration by continuous infusion or twice-daily dosing. Results from these multiple-dose studies indicate that under these conditions of administration, plasma levels of E5564 can be predictive of long-term pharmacodynamic activity.
Immunostimulatory activity of aminoalkyl glucosaminide 4-phosphates (AGPs): induction of protective innate immune responses by RC-524 and RC-529
Authors: Baldridge Jory R. , Cluff Christopher W. , Evans Jay T. , Lacy Michael J. , Stephens Jeffrey R. , Brookshire Valerie G. , Rong Wang  , Ward Jon R. , Yorgensen Yvonne M. , Persing David H. , Johnson David A.
Published: June 2007

Earlier we showed that the structural requirements for adjuvanticity among the aminoalkyl glucosaminide 4-phosphate (AGP) class of synthetic immunostimulants may be less strict than those for other endotoxic activities, including the induction of nitric oxide synthase in murine macrophages and cytokine production in human whole blood. The known role of nitric oxide and pro-inflammatory cytokines in the activation of host defenses against infection prompted us to examine the ability of certain AGPs to enhance non-specific resistance in mice to Listeria monocytogenes and influenza infections as well as to stimulate the production of pro-inflammatory cytokines in mouse splenocytes, human PBMCs, and human U937 histiocytic lymphoma cells. Intranasal administration of RC-524 or RC-529 to mice 2 days prior to a lethal influenza challenge provided significant protection in each case. Similarly, the intravenous administration of these AGPs induced resistance to L. monocytogenes infection as measured by survival or reduction of bacteria in the spleen. Activation of the innate immune response by AGPs appears to involve activation of Toll-like receptor 4 (TLR4) because RC-524 failed to elicit a protective effect in C3H/HeJ mice which have a defect in TLR4 signaling or induce significant cytokine levels in C3H/HeJ splenocytes. Both AGPs also stimulated pro-inflammatory cytokine release in human cell cultures in a dose-dependent manner.
Mal and MyD88: adapter proteins involved in signal transduction by Toll-like receptors
Authors: O'Neill Luke A.J. , Dunne Aisling , Edjeback Michael , Gray Pearl , Jefferies Caroline , Wietek Claudia
Published: June 2007

Signal transduction processes activated by Toll-like receptors (TLRs) include the important transcription factor NF-κB and 2 MAP kinases, p38 and Jun N-terminal kinase. These signals ultimately give rise to increased expression of a multitude of pro-inflammatory proteins. Receptor-proximal proteins involved in signalling by all TLRs include the adapter MyD88, 3 IRAKs (IRAK-4, IRAK and IRAK-2), Tollip, Traf-6 and TAK-1. Differences between signals generated by TLRs are emerging, with both TLR4 and TLR2 signalling requiring an additional adapter termed MyD88-adapter-like (Mal; also known as TIRAP). MyD88 and Mal both have a homologous Toll/IL-1 receptor (TIR) domain although they differ in their N-termini, with MyD88 possessing a death domain. In addition, structural models reveal marked differences in surface charges which, when taken with surface charge differences between TLR2 and TLR4 TIR domains, may indicate that TLR4 but not TLR2 recruits Mal directly. Another difference is that Mal can become phosphorylated. Future studies on Mal will reveal specificities in signal transduction by different TLRs, which may ultimately provide molecular explanations for specificities in the innate immune response to infection.
Differences in the mechanism of nitric oxide production between mouse vascular endothelial cells and macrophages
Authors: Sugiyama Tsuyoshi , Fujita Megumi , Koide Naoki , Morikawa Akiko , Takahashi Kazuko , Yoshida Tomoaki , Mori Hiroshi , Yokochi Takashi
Published: June 2007

The detailed mechanism of NO production in mouse vascular endothelial cells, END-D, was studied. The NO production in END-D cells was triggered by gamma interferon (IFN-γ), but not LPS. However, LPS augmented the NO production in IFN-γ-stimulated END-D cells. A high level of NO production was due to the expression of an inducible type of NO synthase (iNOS) in those cells. A significant amount of NO was detected 18 h after IFN-γ stimulation, accompanied by the delayed iNOS expression. The JAK/STAT signal pathway mediated IFN-γ-induced NO production, but did not participate in the LPS-induced augmentation. Further, no activation of nuclear factor (NF)-κB was involved in the NO production in END-D cells stimulated with either IFN-γ and/or LPS. The mechanism of NO production in END-D cells was suggested to be different from that in mouse macrophages. The differential regulation of NO production in mouse vascular endothelial cells and macrophages is discussed.
The effect of high and low ambient temperatures on PGE2 and TNF-α production by rat glial brain cultures
Authors: Shemi D. , Azab Abed , Kaplanski Jacob
Published: June 2007

This study was undertaken to investigate the effect of changes in incubation temperature on PGE 2 and TNF-α production by rat glial brain cultures after LPS stimulation. One-hour incubation at temperatures of 4°C, 37°C, 39°C and 42°C were used. Treatment of cultures with 10 μg/ml LPS from Escherichia coli caused a significant elevation of PGE2 production 24 h after incubation at control temperatures of 37°C and the experimental temperatures of 4°C, 39°C and 42°C. While high ambient temperatures of 39°C and 42°C reduced LPS-stimulated production, compared to exposure to 37°C, exposure to 4°C did not do so. On the other hand, exposure of the cultures to a temperature of 39°C and 42°C for 1 h did not alter LPS-stimulated TNF-α production, while exposure to a temperature of 4°C significantly reduced this elevation.