bluuebunny/crossref_metadata_2025 huggingface dataset:
Optimisation of pyrogen testing in parenterals according to different pharmacopoeias by probabilistic modelling
Authors: Hoffmann Sebastian , Lüderitz-Püchel Ursula , Montag Thomas , Hartung Thomas
Published: June 2007

The rabbit test to detect pyrogenic contamination in parenterals is crucial to ensure patient safety. The pharmacopoeial tests in Europe, the US and Japan are based on the fever reaction of rabbits, but differ in their experimental design and in their algorithms to assess contamination. Employing an international reference endotoxin, fever can be induced in rabbits. Data from 171 rabbits built the base for probabilistic modelling of the fever reaction and for the comparison of the pharmacopoeial tests. The rabbit fever reaction could be modelled as a function of the amount of injected endotoxin (per kg body weight) by linear regression. Combining the pharmacopoeial algorithms of the rabbit pyrogen test with the developed model allowed analysis of differences regarding test results and animal consumption. This showed that the assessment of pyrogenic contamination strongly depends on the respective pyrogen test stipulated by regulations. Additionally, the approach was used to develop a new experimental design. Two specific versions of this design resulted in a reduction of the number of animals used by about 30% while the safety of the test was maintained. A need for harmonisation is evident, allowing optimisation of the experimental design, which promotes animal welfare.
Crystal structure of a peptidoglycan recognition protein (PGRP) in complex with a muramyl tripeptide from Gram-positive bacteria
Authors: Guan Rongjin , Roychowdury Abhijit , Ember Brian , Kumar Sanjay , Boons Geert-Jan , Mariuzza Roy A.
Published: June 2007

Peptidoglycan recognition proteins (PGRPs) are pattern recognition receptors of the innate immune system that bind, and in some cases hydrolyse, bacterial peptidoglycans (PGNs). We determined the crystal structure of the C-terminal PGN-binding domain of human PGRP-Iα in complex with a muramyl tripeptide representing the conserved core of lysine-type PGNs. The peptide stem of the ligand is buried at the deep end of a long binding groove, with N-acetylmuramic acid situated in the middle of the groove, whose shallow end could accommodate N-acetylglucosamine. Both peptide and glycan moieties are essential for binding by PGRPs. Conservation of key PGN-contacting residues indicates that all PGRPs employ this basic PGN-binding mode. The structure identifies variable residues that likely mediate discrimination between lysine- and diaminopimelic acid-type PGNs. In addition, we propose a mechanism for PGN hydrolysis by Zn2+-containing catalytic PGRPs.
Structure and metabolism of peptidoglycan and molecular requirements allowing its detection by the Drosophila innate immune system
Authors: Mengin-Lecreulx Dominique , Lemaitre Bruno
Published: June 2007

Peptidoglycan (murein) is a major essential and specific constituent of the bacterial cell wall. Its main function is to protect cells against the internal osmotic pressure and to maintain the characteristic cell shape. It also serves as a platform for the anchoring of specific proteins and other cell wall components. This giant macromolecule is composed of long glycan chains cross-linked by short peptides. Any alteration of the disaccharide—peptide basic unit results in a global change of peptidoglycan structure and properties. Such global variations are encountered in nature as conserved variations along phyletic lines but have sometimes been acquired as a result of mutations or as a mechanism of resistance against cell-wall targeted antibiotics. During bacterial cell growth and division, the peptidoglycan mesh is constantly broken down by a set of highly specific hydrolases in a maturation process allowing insertion of newly synthesized units in the pre-existing polymerized material. Depending on the bacterial species considered, degradation fragments are either released in the growth medium or efficiently re-utilized for synthesis of new murein in a sequence of events termed the recycling pathway. Peptidoglycan is one of the main pathogen-associated molecular patterns recognized by the host innate immune system. Variations of the structure and metabolism of this cell wall component have been exploited by host defense mechanisms for detection/identification of invading bacterial species. Modification of the peptidoglycan structure could also represent a mechanism allowing bacteria to escape these host defense systems.
The role of epithelial Toll-like receptor expression in host defense and microbial tolerance
Authors: Hornef Mathias W. , Bogdan Christian
Published: June 2007

The recognition of microbial structures by Toll-like receptors (TLRs) on professional immune cells situated at sterile internal body sites occurs during invasive microbial infection. It indicates infectious non-self and thereby represents the adequate co-stimulatory signal to initiate activation of the adaptive immune system against the invading pathogen. In contrast, most epithelial body surfaces are permanently colonized by microbial organisms of the normal flora and thus TLR ligands are present under physiological conditions. In the following, we discuss the characteristics of TLR-mediated recognition by epithelial cells, the subsequent activation of the host immune system, and protective mechanisms that might help to avoid inadequate stimulation and allow differentiation between commensal or pathogenic micro-organisms. Recent findings suggest that the role of epithelial cells in the maintenance of stable microbial colonization of host surfaces and the immediate host response to infectious challenges might have to be revised.
Lipid-mediated resistance of Gram-negative bacteria against various pore-forming antimicrobial peptides
Authors: Gutsmann Thomas , Hagge Sven O. , David Alexander , Roes Stefanie , Böhling Arne , Hammer Malte U. , Seydel Ulrich
Published: June 2007

Lipopolysaccharides (LPSs) play a dual role as target and as effector molecules. The knowledge of the LPS-induced activation of human immune cells is increasing; however, surprisingly, much less effort seems to be directed towards the understanding of the mechanisms leading to the killing of the bacterial organisms, which eventually results in the release of LPS from the bacterial surface into the blood circulation. We demonstrate mechanisms of interaction of peptides of the innate immune system ( e.g. defensins and cathelicidins) as well as of externally administered antibiotics ( e.g. Polymyxin B) with Gram-negative bacteria. The main focus is directed on data derived from electrical measurements on a reconstitution system of the outer membrane as an asymmetric bilayer composed on one side of LPS and on the other of phospholipids. All these antimicrobial peptides (AMPs) are membrane-active and induce the permeabilization of the reconstituted membranes by the formation of lesions. We found that differences in the activity of the AMPs against various sensitive and resistant Gram-negative bacteria can be explained solely by variations in the chemical structure of LPS, e.g. in the composition of the sugar head group. A reduction of the net negative charge of LPS is responsible for a reduced interaction with the polycationic AMPs and thus for resistance. A most important side effect of positively charged AMPs is the neutralization of the negatively charged LPS released from the bacterial surface as a consequence of AMP-induced killing.
Heritable defects of the human TLR signalling pathways
Authors: Puel Anne , Kun Yang  , Ku Cheng-Lung , von Bernuth Horst , Bustamante Jacinta , Santos Orchidée Filipe , Lawrence Tatiana , Chang Huey-Hsuan , Al-Mousa Hamoud , Picard Capucine , Casanova Jean-Laurent
Published: June 2007

Recently, three human primary immunodeficiencies associated with impaired TLR signalling were described. Anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID), either X-linked recessive or autosomal dominant, is caused by hypomorphic mutations in NEMO or hypermorphic mutation in IKBA, respectively, both involved in nuclear factor-κB (NF-κB) activation. These patients present with abnormal development of ectoderm-derived structures and suffer from a broad spectrum of infectious diseases. In vitro studies of the patients' cells showed an impaired, but not abolished, NF-κB activation in response to a large set of stimuli, including TLR agonists. More recently, patients with autosomal recessive amorphic mutations in IRAK4 have been reported, presenting no developmental defect and a more restricted spectrum of infectious diseases, mostly caused by pyogenic encapsulated bacteria, principally, but not exclusively Gram-positive. In vitro studies carried out with these patients' cells showed a specific impairment of the Toll—interleukin-1 receptor (TIR)—interleukin-1 receptor associated kinase (IRAK) signalling pathway. NF-κB- and mitogen activated protein kinase (MAPK) pathways are impaired in response to all TIR agonists tested. These data, therefore, suggest that TLRs play a critical role in host defence against pyogenic bacteria, but may be dispensable or redundant for immunity to most other infectious agents in humans.
Modulatory effects of sCD14 and LBP on LPS-host cell interactions
Authors: Kitchens Richard L. , Thompson Patricia A.
Published: June 2007

LPS binding protein (LBP) and CD14 play key roles in promoting innate immunity to Gram-negative bacteria by transferring LPS to the signaling receptor complex, MD-2/Toll-like receptor 4 (TLR4). LBP and soluble CD14 (sCD14) can also inhibit responses to LPS by mechanisms that depend on their concentration and environment; during acute inflammation and infection, their concentrations increase in plasma and extravascular fluids. Whereas low concentrations of LBP enhance responses to LPS, high LBP concentrations can inhibit LPS bioactivity in vitro and in vivo. sCD14 also inhibits cell responses by diverting LPS from membrane-bound CD14 (mCD14) and by promoting LPS efflux from cell-surface mCD14 and transferring it to plasma lipoproteins. In vivo studies support the hypothesis that sCD14 has systemic anti-inflammatory effects, whereas in the tissues it may have pro-inflammatory effects that increase resistance to bacteria. Likewise, LBP increases resistance to Gram-negative bacteria by rapidly triggering pro-inflammatory responses to LPS. Thus, the dual stimulatory and inhibitory mechanisms of sCD14 and LBP may benefit the infected host by promoting inflammation in local sites, where it is needed, while at the same time preventing potentially detrimental systemic responses to LPS.
Polymorphism in IL1B: IL1B—511 association with tuberculosis and decreased lipopolysaccharide-induced IL-1β in IFN-γ primed ex-vivo whole blood assay
Authors: Awomoyi Agnes A. , Charurat Manhattan , Marchant Arnaud , Miller E. Nancy , Blackwell Jenefer M. , McAdam Keith P.W.J. , Newport Melanie J.
Published: June 2007

To determine whether variation in two interleukin 1 family genes ( IL1B and interleukin 1 receptor antagonist, IL1RN) is associated with pulmonary tuberculosis (TB), two published polymorphisms at nucleotide positions —511 and +3953 in IL1B and one in the IL1RN 86 bp VNTR were genotyped in 335 smear positive Gambian TB patients, and 298 ethnically matched controls. All individuals were HIV negative. Decreased risk of pulmonary TB was associated with both heterozygosity and homozygosity for the IL1B—511-C allele (OR 0.66, P = 0.027, and OR 0.58, P = 0.015, respectively). Nonetheless, the C allele was present at a frequency of 0.66 in TB cases suggesting that whilst IL-1β contributes to disease susceptibility, it is not the major factor. There was no association between the IL1B+3953-T/C polymorphism or the 86 bp IL1RN pentallelic repeat and TB in this population. Using an ex-vivo whole blood assay, healthy Gambian individuals who are homozygous for the IL1B—511-T allele failed to exhibit a significant increase in IL-1β production in response to LPS after IFN-ypriming.
Conserved mechanisms of signal transduction by Toll and Toll-like receptors
Authors: Gangloff Monique , Weber Alexander N.R. , Gay Nicholas J.
Published: June 2007

In recent years, considerable progress has been made towards understanding the mechanism by which endotoxin is detected by the cells of the immune system. Lipopolysaccharides are extracted in a soluble form by the serum LPS binding protein and then transferred sequentially to the extrinsic membrane protein CD14 and the co-receptor complex TLR4/MD-2. Our modelling studies suggest that acyl chains of lipid A are buried within the hydrophobic core of MD-2 and this induces crosslinking of the two TLR4/MD-2 complexes, an event that is required to trigger signal transduction. We also propose that, by analogy with the Drosophila Toll receptor, the mechanism of signal transduction is likely to be complex and to involve concerted protein conformational changes. In particular, we propose that receptor—receptor interactions mediated by juxtamembrane sequences play a critical role.
How Toll-like receptors and Nod-like receptors contribute to innate immunity in mammals
Authors: Fritz Jörg H. , Girardin Stephen E.
Published: June 2007

Innate immune detection of pathogens relies on specific classes of microbial sensors called pattern-recognition molecules (PRM). In mammals, such PRM include Toll-like receptors (TLRs) and the intracellular proteins NOD1 and NOD2, which belong to the family of Nod-like receptors (NLRs). Over the last decade as these molecules were discovered, a function in innate immunity has been assigned for the majority of them and, for most, the microbial motifs that these molecules detect were identified. One of the next challenges in innate immunity is to establish a better understanding of the complex interplay between signaling pathways induced simultaneously by distinct PRMs and how this affects tailoring first-line responses and the induction of adaptive immunity to a given pathogen.
Antibodies against human Toll-like receptors (TLRs): TLR distribution and localization in human dendritic cells
Authors: Seya Tsukasa , Funami Kenji , Taniguchi Mitsue , Matsumoto Misako
Published: June 2007

We have produced monoclonal antibodies (mAbs) against human Toll-like receptors (TLRs). These mAbs recognize the natural conformation of the extracellular domain leucine-rich repeats and, thereby, are suitable for immunoprecipitation, flow cytometric analysis and immunostaining. Using these mAbs, we determined the distribution of TLRs in a variety of human cell populations. Human TLRs that recognize bacterial components, particularly TLR-1, TLR-2, TLR-4 and TLR-6, reside on the cell surface and are expressed in myeloid and monocyte-derived dendritic cells (DCs) but not in plasmacytoid DCs (pDCs). Human TLR-3 resides in putative endosomes in myeloid DCs. Thus, the human myeloid DC subset harbors a unique and distinctive TLR repertoire. These mAbs will be useful to test the localization and distribution of human TLRs in a variety of cells and organs. Functional studies of human TLRs will also be feasible since certain of them are function-blocking mAbs.
Invited review: Origin of monocytes and their differentiation to macrophages and dendritic cells
Authors: Kumar Suresh , Jack Robert
Published: June 2007

Activation of the immune system requires that the presence of an incipient infection first be detected. This essential step is carried out by tissue macrophages, which alert innate immunity, and by dendritic cells whih alert the adaptive immune system. Both of these sentinel cell types are derived from circulating precursors known as blood monocytes. Here we briefly review current work which has recently expanded our understanding of the origin of blood monocytes and of the factors governing their further differentiation into tissue sentinels.
Effects of human apolipoprotein A-I on endotoxin-induced leukocyte adhesion on endothelial cells in vivo and on the growth of Escherichia coli in vitro
Authors: Thaveeratitham Premtip , Plengpanich Wanee , Naen-Udorn Worakamol , Patumraj Suthiluk , Khovidhunkit Weerapan
Published: June 2007

Background: High-density lipoprotein (HDL) has been shown to inhibit leukocyte adhesion to endothelial cells induced by endotoxin in vivo and suppress the growth of bacteria in vitro; however, the components responsible for these effects, either lipids or proteins, are not yet defined. In this study, we examined the effects of apolipoprotein (apo) A-I, the major protein of HDL, on ameliorating the effect of endotoxin and inhibiting the growth of bacteria.

Materials and Methods: Apo A-I, purified from normal human HDL, was incubated with endotoxin. Leukocyte adhesion to endothelial cells of rat mesenteric venules was assessed using intravital fluorescence microscopy. Ability of apo A-I to inhibit the growth of Escherichia coli was assessed using a spread plate method.

Results: Purified, lipid-free apo A-I could inhibit endotoxin-induced leukocyte adhesion to endothelial cells in vivo in a dose-dependent manner. In addition, apoA-I was able to suppress the growth of Escherichia coli in vitro.

Conclusions: These data suggest that apo A-I of HDL can directly interact with endotoxin, ameliorating its effect and that apo A-I may have a direct toxic effect on whole bacteria. Therefore, therapeutic use of apo A-I in septicemia and bacterial infection should be further explored.
Inflammatory responses associated with acute coronary syndrome up-regulate IRAK-M and induce endotoxin tolerance in circulating monocytes
Authors: del Fresno Carlos , Soler-Rangel Llanos , Soares-Schanoski Alessandra , Gómez-Piña Vanesa , González-León María Carmen , Gómez-García Lourdes , Mendoza-Barberá Elena , Rodríguez-Rojas Alexandro , García Felipe , Fuentes-Prior Pablo , Arnalich Francisco , López-Collazo Eduardo
Published: June 2007

Acute coronary syndrome (ACS) groups different cardiac diseases whose development is associated with inflammation. Here we have analyzed the levels of inflammatory cytokines and of members of the TLR/IRAK pathway including IRAK-M in monocytes from ACS patients classified as either UA (unstable angina), STEMI (ST-elevation myocardial infarction) or NSTEMI (non-ST-elevation myocardial infarction). Circulating monocytes from all patients, but not from healthy individuals, showed high levels of pro-inflammatory cytokines, TNF-α and IL-6, as well as of IRAK-M and IL-10. TLR4 was also up-regulated, but IRAK-1, IRAK-4 and MyD88 levels were similar in patients and controls. Further, we investigated the consequences of cytokines/IRAK-M expression on the innate immune response to endotoxin. Ex vivo responses to LPS were markedly attenuated in patient monocytes compared to controls. Control monocytes cultured for 6 h in supplemented medium (10% serum from ACS patients) expressed IRAK-M, and LPS stimulation failed to induce TNF-α and IL-6 in these cultures. Pre-incubation of the serum with a blocking anti-TNF-α antibody reduced this endotoxin tolerance effect, suggesting that TNF-α controls this phenomenon, at least partially. We show for the first time that inflammatory responses associated with ACS induce an unresponsiveness state to endotoxin challenge in circulating monocytes, which correlates with expression of IRAK-M, TLR4 and IL-10. The magnitude of this response varies according to the clinical condition (UA, STEMI or NSTEMI), and is regulated by TNF-α.
Involvement of Syk protein tyrosine kinase in LPS-induced responses in macrophages
Authors: Ulanova Marina , Asfaha Samuel , Stenton Grant , Lint Alison , Gilbertson Deanna , Schreiber Alan , Befus Dean
Published: June 2007

Syk kinase is best known as a critical component of immunoreceptor signaling in leukocytes. Activation of Syk following cross-linking of Fcγ and Fcε receptors on macrophages, mast cells, and other cells induces various inflammatory events. We hypothesized that Syk is involved in inflammatory responses induced by the lipopolysaccharide (LPS). We studied the role of Syk using its inhibition by antisense oligonucleotides, or small interfering RNA. Our data demonstrated that in vivo inhibition of Syk caused down-regulation of LPS-induced responses in rat alveolar macrophages. In in vitro experiments, inhibition of Syk in rat peritoneal macrophages, as well as in human myelomonocyte cell line THP-1 also caused a decrease in LPS-induced cytokine release. Our data support the hypothesis that, in macrophages, Syk is involved in LPS-induced intracellular signaling pathways leading to the release of pro-inflammatory mediators. Understanding the role of Syk in LPS-induced signaling may help in developing new therapeutic tools for inflammatory disorders.
Cell surface binding of LBP—LPS complexes to a protein component distinct from CD14
Authors: Tapping Richard I. , Tobias Peter S.
Published: June 2007

Lipopolysaccharide (LPS) binding protein (LBP) is a serum molecule that mediates cellular activation in response to endotoxin by ensuring the delivery of LPS to either soluble or membrane bound forms of CD14. Aside from this activating role, previous work has shown that LBP and LPS can bind to cells by forming large aggregates which are anchored by mCD14. This binding phenomenon does not correlate with cellular activation. To further characterize these events, we have generated a biologically active radiolabeled LBP ligand with high specific activity. Through the use of this ligand in whole cell binding assays, we have confirmed that the binding of LBP to CHO cells expressing mCD14 is LPS dependent, blocked by the anti-LBP antibodies 18G4 and 2B5, and appears to involve the self aggregation of LBP—LPS complexes on the cell surface. Moreover, we discovered that non-transfected CHO cells also exhibit a binding phenomenon with all the above characteristics of CHO-mCD14 cells. Binding through this latter receptor(s) is distinct from that mediated by mCD14 in that it is not inhibited by anti-CD14 antibodies 28C5 or 18E12. In addition, unlike binding to mCD14, binding of LBP—LPS complexes to this novel receptor is abolished by pretreatment of cells with trypsin. Using proteinase K we found that LBP—LPS complexes bound either by mCD14 or this new receptor are subsequently internalized. Pretreatment of cells with trypsin also abolishes their ability to internalize mCD14 bound LBP—LPS complexes. The novel receptor for LBP—LPS complexes has been detected on many cell types and may be a receptor required for the cellular clearance of LPS.
Biosynthesis and genetics of ADP-heptose
Authors: Valvano Miguel A.
Published: June 2007

Glycero- manno-heptose is a common component in the lipopolysaccharide (LPS) of many Gram-negative bacteria. Mutants deficient in the synthesis of glycero- manno-heptose are highly sensitive to hydrophobic compounds, and display reduced virulence, making these genes and their products potential targets for developing novel antimicrobials. To date, the biosynthesis of the heptosyl precursors for the inner core oligosaccharide of the LPS molecule is not completely characterized. In this work, the genes and enzyme functions involved in the various steps of the biosynthesis of ADP-L- glycero-D- manno-heptose are discussed, especially those involved in the intermediate steps.
Glycosyltransferases of Pseudomonas aeruginosa that assemble the O antigens of A band and B band lipopolysaccharide
Authors: Lam J.S. , Rocchetta H.L. , Burrows L.L.
Published: June 2007

Pseudomonas aeruginosa produces two forms of lipopolysaccharide (LPS) designated A band and B band. The O-polysaccharide region of A band is a conserved D-rhamnan polymer arranged α1—2, α1—3, α1—3, while B band is serotype-specific with differences in the O-antigenic region dividing P. aeruginosa into 20 stereotypes. The B band O-antigen unit of serotype O5 is [—4)-β-D-Man(2NAc3N)A-(1-4)-β-D-Man(2NAc3NAc)A-(1-3)-α-D-Fuc2NAc]. The glycosidic structure of LPS molecules specified by the action of dedicated glycosyltransferases. The wbp clusters of A band and B band (serotype O5) were each found to contain three genes coding for putative glycosyltransferases: wbpX, wbpY, wbpZ, and wbpH, wbpJ, wbpL , respectively. To examine the role of these potential transferases in LPS assembly, chromosomal mutations were generated within all 6 genes. LPS analysis reveals that wbpX, wbpY, and wbpZ mutants express an A—B+ phenotype, while wbpH and wbpJ mutants are A+B—. Interestingly, mutations in wbpL, an Escherichia coli wec A homologue, abrogates both A band and B band LPS synthesis. Based on amino acid homologies, O-polysaccharide structures and LPS phenotypes of transferase mutants, we propose an assembly scheme for these two LPS molecules.
Lipopolysaccharide- and paclitaxel (Taxol)-induced tolerance in murine peritoneal macrophages
Authors: Nakano Masayasu , Tominaga Kaoru , Saito Sinji , Kirikae Fumiko , Songnian Lin  , Fumero Cecilia L. , Ojima Iwao , Kirikae Teruo
Published: June 2007

LPS-tolerance, a state of refractoriness to LPS-stimulation, is induced in murine peritoneal macrophages by prior exposure to LPS. LPS-induced expression of TNF and IL-6 mRNA as well as activation of various intracellular kinases and factors, including ERK, p38, JNK, Raf-1 and NF-κB were all suppressed in LPS-tolerant macrophages; responses to stimulation by paclitaxel (Taxol™), an LPS agonist, were similarly suppressed, but responses to phorbol esters (PMA) were unaffected. Binding and uptake of [125I]-labeled LPS to tolerant macrophages was somewhat greater in tolerant than in non-tolerant macrophages. Thus, the refractory state appears to involve inhibition or blockade of LPS-signaling molecules located downstream of the cell membrane LPS receptor and upstream of the branch point in the intracellular cascades leading to activation of MAPK and NFκB. LPS conditioning also suppressed LPS- and Taxol-induced TNF production, but augmented nitric oxide (NO) production. In contrast, Taxol conditioning failed to suppress LPS-induced TNF production. Conditioning with the synthetic taxoid analog, nor-seco-taxoid, which does not induce macrophage activation, enhanced LPS- and Taxol-induced NO production. These findings provide us with new information about the relationship between the LPS and Taxol receptors as well as about the signaling pathways leading to TNF and NO production.
Activation of protein kinase CK2 by LPS is mediated by the MAP kinase pathway
Authors: Means Terry K. , Lodie Tracey A. , Fenton Matthew J.
Published: June 2007

Stimulation of macrophages by Gram-negative bacterial lipopolysaccharide (LPS) rapidly leads to the activation of several protein kinases and the subsequent phosphorylation of transcription factors that regulate LPS-inducible genes. Several investigators have shown that the MAP kinases (MAPKs) are rapidly activated following LPS stimulation. We previously reported that LPS stimulation also up-regulates the enzymatic activity of protein kinase CK2, a ubiquitous serine/threonine kinase, although the signaling cascade that leads to CK2 activation in macrophages is unknown. Because MAPKs are known to be rapidly activated by LPS, we performed additional studies in order to determine if CK2 activation was a downstream target of MAPKs. Our studies revealed that CK2 activity was rapidly and transiently up-regulated in LPS-stimulated RAW264.7 murine macrophages. We found that PD98059, an inhibitor of ERK kinase activation by the MAP kinase MEK-1, blocked LPS-induced up-regulation of CK2 activity. In contrast, the p38 kinase inhibitor SB203580 did not block CK2 activation by LPS. This finding suggests that CK2 activation was mediated by the ERK kinase signaling cascade, but not by the p38 kinase cascade. We also found that LPS stimulation resulted in the rapid serine phosphorylation of the catalytic α and α′ subunits of CK2. This contrasts with other studies using growth factor-stimulated fibroblasts in which only phosphorylation of the regulatory β subunit of CK2 was observed. Thus, LPS stimulation leads to rapid activation and cell type-specific phosphorylation of CK2 in macrophages.
New synthesis and conformational analysis of lipid A: biological activity and supramolecular assembly
Authors: Fukase Koichi , Oikawa Masato , Suda Yasuo , Liu Wen-Chi , Fukase Yoshiyuki , Shintaku Tetsuya , Sekljic Harald , Yoshizaki Hiroaki , Kusumoto Shoichi
Published: June 2007

In order to investigate precise structural requirements for expression of the biological activity of lipid A, new structural analogs were synthesized via improved, highly efficient preparative routes. The number and chain length of the fatty acyl groups proved to have crucial influence on the biological activity. For the study of conformation relevant to activity of lipid A by means of NMR spectrometry, lipid A analogs [13C]-labeled at the 6-position were then synthesized starting from 6-[13C]-glucose. Precise analysis of J CH and NOE data disclosed a particular molecular conformation and characteristic supramolecular assembly in an aqueous SDS micelle solution of the tetraacyl biosynthetic precursor molecule. The observed mode of assembly may reflect the actual behavior of lipid A molecules in the cell surface outer membrane of living bacteria. A biologically inactive, artificial, tetra-acyl analog having shorter acyl chains exhibits, by contrast, distinct conformations with no sign of supramolecule formation. The acyl moieties in lipid A are thus found to play an important role in regulating the overall conformation of the hydrophilic backbone.
Identification of CD18 as a novel Taxol binding/signaling protein in murine macrophage membranes
Authors: Vogel Stefanie N. , Bhat Nayantara , Carboni Joan M. , Mayadas Tanya N. , Blanco Jorge , Perera Pin-Yu , Golenbock Douglas T.
Published: June 2007

The anti-tumor agent, Taxol, is a potent LPS mimetic in murine macrophages, an activity that is dissociable from its well-characterized anti-mitotic activity which is mediated by microtubule hyperstabilization. A photoactivatable Taxol analog was used to identify components of a putative, shared LPS signaling apparatus in murine macrophage membranes. We report here that CD18, the β chain of the β2-integrin, Mac-1, represents a major Taxol binding protein in murine macrophages.
Structure of lipid A and cell activation
Authors: Qureshi Nilofer , Kutuzova Galina , Takayama Kuni , Rice Peter A. , Golenbock Douglas T.
Published: June 2007

Lipopolysaccharide (LPS) is the principal antigen of Gram-negative bacteria. The free lipid A is derived by mild acid hydrolysis of the LPS. The structures of lipid A from three select sources were compared. The toxic lipid A of Enterobacteriaceae LPS contains 6 fatty acids (C14 or C12) whereas the nontoxic penta-acyl diphosphoryl lipid A derived from the LPS of Rhodobacter sphaeroides (RsDPLA) contains short-chain fatty acids (C10). The nontoxic penta-acyl lipid A from Chlamydia trachomatis contains long-chain fatty acids (C20). This analysis shows that the toxic property of lipid A is determined by the fatty acid composition. Clearly, there is a fine structural requirement for toxicity (and biological activities) of lipid A (including LPS). As an effective antagonist in both human and murine cell lines, RsDPLA is a useful reagent for studying toxic LPS-induced signaling. RsDPLA appears to bind to the putative physiological receptor and does not allow the toxic LPS to bind and initiate signaling. This appears to occur very early at the level of both LBP and CD14. This suggests that RsDPLA may be a useful drug for Gram-negative septic shock.
The Effectiveness of a Psychoeducational Group for HIV-Infected/Affected Incarcerated Women
Authors: Pomeroy Elizabeth C. , Kiam Risa , Abel Eileen M.
Published: June 2007

Objective: The effectiveness of a psychoeducational group intervention for HIV/AIDS-infected and affected women was examined at a large southeastern county jail facility. Method: A quasiexperimental pretest-posttest design was used to examine depression, anxiety, and trauma symptoms of women inmates. Results: A multivariate analysis of covariance yielded significant differences between the experimental and comparison groups. Subsequent analysis of covariance for each dependent variable indicated significant differences between groups as well. Effect sizes ranged from moderate to strong. Conclusions: The psychoeducational group intervention appeared to be effective in alleviating depression, anxiety, and trauma symptoms among women inmates infected and affected by HIV/AIDS.
Placement Restrictiveness and Educational Achievement Among Emancipated Foster Youth
Authors: Mech Edmund V. , Fung Carrie Che-Man
Published: June 2007

Objective: To investigate the relationship between measures of placement restrictiveness and educational achievement for a sample of emancipated foster youth. Method: Information was collected on 171 foster adolescents in Illinois. The permanency plan for each ward was independent living. Three methods were used to calculate the cumulative restrictiveness levels of various living arrangements while in placement. Follow-up information on educational progress was collected at age 21. Results: Two thirds of the youths in less restrictive placements attended postsecondary education, compared to one third who were placed in highly restrictive settings. Overall, nearly 85% of the enrollees in postsecondary education or training programs came from placements rated as low in restrictiveness. Conclusions: Placements rated as low in restrictiveness such as foster family homes and transitional apartments are probably the most effective settings in which to prepare foster wards for independence. Overall, the most useful method for calculating restrictiveness was the Predominant Placement Type formula.
The Centre for Child Care Research at the Queen's University of Belfast
Authors: Iwaniec Dorota , McCrystal Patrick
Published: June 2007

The Centre for Child Care Research at the Department of Social Work, The Queen's University of Belfast, was established in 1995 through a partnership between the Department of Health and Social Services, the four Health and Social Services Boards, and the Social Work Department at The Queen's University. The Centre was funded to play a key role in influencing the development of child care policy and practice in Northern Ireland, as well as to link its research to national and international levels following the introduction of major child care legislation, the Children (Northern Ireland) Order 1995. This article examines the value of research centers in developing policies and evidence-based practice generally, but specifically focuses on the Centre for Child Care Research, its research program, organizational structures, and other activities.
How do surrogate decision makers describe hospice? Does it matter?
Authors: Vig Elizabeth K. , Starks Helene , Taylor Janelle S. , Hopley Elizabeth K. , Fryer-Edwards Kelly , Pearlman Robert A.
Published: June 2007

We interviewed 71 surrogate decision makers of older, chronically ill veterans to explore their knowledge of hospice and their role in helping loved ones access hospice services. We asked them to describe hospice and any previous hospice experiences. Qf the group, 24 percent with hospice experience and 14 percent without hospice experience correctly described three key aspects of hospice: who hospice cares for, where the care is provided, and the goal of the care. Additionally, we found evidence that surrogates who correctly described the three key aspects of hospice were inclined to pursue hospice care for loved ones in the future, and surrogates who provided less complete descriptions of hospice might not access it. Since surrogates often help dying patients access care, incomplete knowledge of hospice may be an important barrier to hospice services. We advocate that clinicians discuss the three key aspects of hospice during routine advance care planning sessions with patients and their future surrogate decision makers.
Who's caring for whom? Differing perspectives between seriously ill patients and their family caregivers
Authors: Hauser Joshua M. , Chang Chih-Hung , Alpert Hillel , Baldwin DeWitt , Emanuel Ezekiel J. , Emanuel Linda
Published: June 2007

Although clinicians and researchers often rely on family members' reports of a wide range of dying patients' symptoms and care preferences, available data indicate divergences between the two. We used a national sample to analyze patient-caregiver pairs to explore areas of concordance and nonconcordance about physical symptoms, communication with physicians, caregiving needs, andfuturefears. We also assessed whether identifiable patient or caregiver characteristics were associated with nonconcordance. Our data were from a national, random sample of 988 terminally ill patients, of whom 893 had caregivers who were also interviewed. Frequencies and types of nonconcordance were computed for patient-caregiverpairs. Bivariate associations between patient and caregiver reports on each item were tested Logistic and conditional logistic regression analyses assessed multiple predictors of nonconcordance for each item. Primary diagnoses included cancer (51.1 percent), heart disease (17. 7 percent), chronic obstructive pulmonary disease (10. 7percent), and other diseases (20.5 percent). The proportion of concordant reports among pairs ofpatients and caregivers ranged from 53 percent to 66 percent. Among pairs showing nonconcordant responses, caregivers reported higher levels of pain and disability than patients, lower caregiving needs, and different fears about thefuture. Few demographic or clinical predictors were associated with nonconcordance. Concordance between patients' and their caregivers' responses ranged widely, and there were important areas of nonconcordant responses. When responses differed, patients were more likely to express concern about domains that might impose on caregivers, while caregivers were more likely to express concern about patients' physical suffering. Consistent sociodemographic or clinical predictors of nonconcordant responses were not found These data suggest important ways that patient and caregiver reports of the samg e experience vary.
Antidepressant medication use in palliative care
Authors: Miller Karl E. , Adams Stephen M. , Miller Martha M.
Published: June 2007

Depression and other mood disorders occur in approximately 25 percent of terminal patients. Untreated, depression and mood disorders can have a significantly negative impact on patients andfamilies. Screeningfor depression can be done as easily as asking one question: “Areyou depressed?” A positive response to this question can be followed with one of the more extensive screening tools. Anxiety disorders can also have a negative effect on patients and their families. These can be identified by also using one of the validated screening tools. Use of the antidepressant medications for treating depression and, in some cases, anxiety disorders has not been well studied in hospice and palliative care. Some of the antidepressants can also serve as adjuvant therapy in pain management.
Responding to the needs of the terminally ill through laughter and play
Authors: Graham Linda L. , Cates Jim A.
Published: June 2007

Carefor the term inally ill has greatly expanded. Humor and play, however, remain largely unexplored, surrendered to the cultural expectation of dignity and respect for the dying. This paper suggests several uses of humor and play with the dying, and the benefits of these interventions with patients, families, and care givers.
The effects of death education on nurses' attitudes toward caring for terminally ill persons and their families
Authors: Frommelt Katherine H. Murray
Published: June 2007

This study sought to determine the cffectiveness of an education program on nurses' attitudes toward caring for terminally ill persons and their family members. The program, based on the hospice concept of care, included a didactic section based on Kubler-Ross' stages of death and dying, and a roleplay model designed by the researcher. Data were collected from 34 licensed nurses, aged 18 to 65, practicing in the midwestern United States. The Frommelt Attitude Toward Care of the Dying Scale (FATCOD) was designed by the researcher to assess nurses' attitudes. The FATCOD was found to be a valid and reliable tool. All nurses completed the tool before and after the education program (pre-test, post-test). Compared by a t-test, the scores for the nurses were sign Qflcantly higher after participation in the educational program. The t-value wasfound to be 2.97, significant at the < 0.01 level, 2-tailed probability = 0.006. These findings support the hypothesis that nurses have a more positive attitude toward caring for terminally ill persons and their family members after participation in the program, than the same nurses had before participating in the program. Demographic information including age, years of experience in nursing, highest degree held, basic type of nursing preparation and previous education on death and dying were analyzed to determine their relationship to the nurses' attitudes. The only information which demonstrated any significant relationship to the nurses' attitudes was that ofprevious education on death and dying. These were computed by an analysis of variance (ANOVA) F = 3.22, F prob = 0.04, significant at < 0.05 level. Of the nurses who participated in the study, 76.5 percent indicated that they were unhappy with the education which they hadpreviously received on the subject of death and dying.
Prospective trials for the radiotherapeutic treatment of bone metastases
Authors: Berk Lawrence
Published: June 2007

Radiation therapy is a proven technique to relieve the pain of bone metastases. Focal painful metastases can be palliated in up to 80 percent of patients. The technique to use for the treatment varies depending on the extent of disease, the prognosis of the patient and physician preference. Most studies indicate that the local treatment of bone metastases with single large treatments is as efficacious as extended courses of treatment. For patients with widespread disease, two forms of systemic radiotherapy are available: hemibody irradiation and intravenous injection of radionuclides. Studies have shown the combination of either focal irradiation and hemibody irradiation or focal irradiation and the injection of strontium(89) prolongs the pain-free duration of the patients. All palliative patients with symptomatic bone metastases should be evaluated for radiotherapy.
Implementation and evaluation of a quality improvement process to improve pain management in a hospice setting
Authors: Duggleby Wendy , Alden Christine
Published: June 2007

The purpose of this article is to describe the implementation and evaluation of a quality improvement process to improve pain management in a hospice setting. A retrospective chart audit of 702 patient visits pre- and 536 patient visits post-implementation of quality improvement strategies measured five aspects of pain management: complaints of pain, severity of pain, changes in patient’s pain medication regime required, patient and family teaching, and use of complementary therapies. Of these measures a significant change was found in the documentation of pain assessment, recognition of changes required in the medication regime, and patient and family teaching.
Multisystemic Therapy: An Empirically Supported, Home-Based Family Therapy Approach
Authors: Sheidow Ashli J. , Woodford Mark S.
Published: June 2007

Multisystemic Therapy (MST) is a well-validated, evidenced-based treatment for serious clinical problems presented by adolescents and their families. Across randomized clinical trials, MST has been effective in reducing out-of-home placements, delinquent behavior, substance use, and psychiatric symptomatology compared to services currently available in the community. This article is an introduction to the MST approach and outlines key clinical features, describes the theoretical underpinnings, and discusses the empirical support for MST’s effectiveness with a variety of serious clinical issues.
Ethical Considerations of Family Secret Disclosure and Post-Session Safety Management
Authors: Fall Kevin A. , Lyons Christy
Published: June 2007

The ethical issues involved in the disclosure of family secrets in therapy have been addressed in the literature, but the focus has typically been on secrets disclosed in individual sessions. The literature largely ignores the ethical issues surrounding in-session disclosure and the concomitant liability of the family therapist for the post-session well-being of the system’s members. This article explores types of family secrets, provides a case example of in-session disclosure, and presents ethical considerations and practice recommendations.
Wait Not, Want Not: Factors Contributing to the Development of Anorexia Nervosa and Bulimia Nervosa
Authors: Murray Trish
Published: June 2007

Eating disorders have grown to epidemic proportions in our culture. The purpose of this literature review is to examine prevalence and incident rates of both anorexia nervosa and bulimia nervosa. In addition, this article will review the psychological and sociological factors that contribute to the development and maintenance of an eating disorder. Finally, different treatment approaches will be discussed in relation to their effectiveness in addressing anorexia or bulimia.
The Ethical Genogram: A Tool for Helping Therapists Understand their Ethical Decision-Making Styles
Authors: Peluso Paul R.
Published: June 2007

The purpose of this article is to address the underlying emotional factors that contribute to therapists’ difficulties in making decisions in accordance with ethical codes as well as introduce a mechanism for assessing the family-of-origin dynamics that shape therapists’ ethical decision-making processes. The ethical genogram can allow therapists to gain insights into the emotionally driven forces that can lead to difficulty making tough ethical decisions. Using the ethical genogram may help guide supervisees as they wrestle with ethical questions and develop their ethical decision-making skills. Suggestions for using the ethical genogram in training and supervision are included.
Live Supervision: A Review of the Research
Authors: Champe Julia , Kleist David M.
Published: June 2007

This article reviews research on live supervision published within the past 12 years. The focus of the research on live supervision can be conceptualized as addressing its prevalence and use, the impact of live supervision interventions, and perceptions of live supervision by supervisors, trainees, and clients. Implications for supervisors, practitioners, and future research are discussed.
Dysplasia in Chronic Ulcerative Colitis: A Molecular Approach to Its Differential Diagnosis
Authors: Urbanski Stefan J. , Fogt Franz
Published: June 2007

Ulcerative colitis-related epithelial dysplasia represents a premalignant lesion, which may lead to the development of ulcerative colitis-related adenocarcinoma. The proper management of this condition requires proctocolectomy before the acquisition of the invasive phenotype. Patients with chronic ulcerative colitis, however, like the general population, may develop sporadic colorectal adenomas not related to the presence of the inflammatory bowel disease. These adenomas are also characterized by epithelial dysplasia, but their detection carries very different clinical implications. It is therefore essential to distinguish between chronic ulcerative colitis-related dysplasia and sporadic colorectal adenomas. The gross and histologic features do not provide a definite distinction between these two different types of dysplasia of the colonic epithelium. Recent developments in the molecular genetics may provide the necessary means.