bluuebunny/crossref_metadata_2025 huggingface dataset:
Pro‐inflammatory cytokines induce c‐fos expression followed by IL‐6 release in human airway smooth muscle cells
Authors: McKay Sue , Bromhaar Mechteld M. Grootte , de Jongste Johan C. , Hoogsteden Henk C. , Saxena Pramod R. , Sharma Hari S.
Published: November 2003

Background: Airway smooth muscle (ASM) is considered to be a target for mediators released during airway inflammation.

Aims: To investigate the expression of c‐fos, a constituent of the transcription factor activator protein‐1, in human ASM cells. In addition, to measure the release of interleukin (IL)‐6 into the conditioned medium of stimulated ASM cells, as well as DNA biosynthesis and changes in cell number.

Methods: Serum‐deprived human ASM cells in the G0/G1 phase were stimulated with the pro‐inflammatory cytokines; tumour necrosis factor‐α, IL‐1β, IL‐5 and IL‐6. The expression of mRNA encoding the proto‐oncogene c‐fos was measured by Northern blot analysis. Cell proliferation was assessed by
{3H}‐thymidine incorporation assays and cell counting, and IL‐6 levels in cell‐conditioned medium were measured by enzyme‐linked immunosorbent assay.

Results: All of the cytokines investigated induced a rapid (within 1h) and transient increase in the expression of mRNA encoding c‐fos, followed by the expression and enhanced release of IL‐6. Cell proliferation remained unchanged in cytokine‐stimulated cells.

Conclusions: Cytokine‐induced c‐fos expression in human ASM cells could be described as a marker of cell ‘activation′. The possible association of these results with airway inflammation, through secondary intracellular mechanisms such as cytokine production, is discussed.
Competition of IL‐1 and IL‐1ra determines lymphocyte response to delayed stimulation with PHA
Authors: Dąbrowski Marek P. , Stankiewicz Wanda , Płusa Tadeusz , Chciałowski Andrzej , Szmigielski Stanisław
Published: November 2003

Background: Human peripheral blood mononuclear cells (PBMC) left in microcultures for 24 h without mitogen do not respond to subsequent stimulation with PHA. They regain reactivity if the native culture medium is absorbed with other party lymphocytes or partially replaced with the medium from a PHA‐stimulated culture. The observations suggest that, during the incubation, some inhibitory agent had accumulated in the culture medium.

Aim: The study was performed to determine the nature of the observed phenomenon in respect of the possible role of monocytes and their products IL‐1 and IL‐1 receptor antagonist (IL‐1ra), and to test for immunodiagnostic purposes the significance of quantifying the lymphocyte response to delayed stimulation with PHA in patients suffering from inflammatory prosesses.

Methods: Lymphocyte response to delayed stimulation with PHA, calculated as the lymphocyte‐monokine interaction (LM) index, was determined in the microcultures of PBMC isolated from the blood of healthy donors or of patients with acute tonsilitis. The values of LM indices were compared with the ratios of IL‐1ra/IL‐1β concentration estimated by enzyme‐linked immunosorbent assay method in the culture supernatants. The influences of exogenous IL‐1β , IL‐1ra, anti‐IL1ra antibodies and antibiotic cefaclor on the monokine concentrations and on the values of LM index were tested.

Results and conclusions: The results show that the level of lymphocyte response to delayed stimulation with PHA (LM index) is inversely proportional to the ratio of IL‐1ra/IL‐1β concentration in the culture. The low LM values at high IL‐1ra/IL‐1β ratios in PBMC cultures from healthy donors, reversed proportions found in patients′ PBMC (acute tonsilitis), and the cefaclor‐induced reduction of LM value with correlated increase of the IL‐1ra/IL‐1β ratio suggest that the LM assay may prove to be useful for immunodiagnostic purposes.
The presence of elafin, SLPI, IL1‐RA and STNFα RI in head and neck squamous cell carcinomas and their relation to the degree of tumour differentiation
Authors: Westin U. , Nyström M. , Ljungcrantz I. , Eriksson B. , Ohlsson K.
Published: November 2003

Biopsy samples of head and neck carcinomas were investigated with regard to elafin, secretory leukocyte protease inhibitor (SLPI), interleukin 1‐receptor antagonist [(IL)1‐RA] and soluble tumour necrosis factor α receptor antagonist (STNFα RI). SLPI and elafin are serine protease inhibitors produced in the serous cells of the upper respiratory airways and in the keratinocytes, respectively. We have now found the presence of elafin and SLPI in squamous cell carcinomas of the upper respiratory tract (tonsillar, hypopharyngeal, tongue, mouth floor, gingival and laryngeal cancer). Significantly higher amounts of SLPI and elafin are present in well‐differentiated and moderately differentiated tumours than in poorly differentiated tumours (p < 0.0001
and p < 0.0015).
Tumour necrosis factor‐α and IL‐1β have been shown to stimulate the production of SLPI and elafin. Since these cytokines can both be difficult to detect, we chose to study their inhibitors, STNF α RI and IL1‐RA, instead. IL1‐RA was expressed in highly differentiated tumours as well as in poorly differentiated ones. No significant difference was seen between the groups. STNF α RI was only found in very small amounts, sparsely distributed in the tumours, and was not related to the degree of differentiation.
Pharmacological characterisation of arthritis induced by Bothrops jararaca venom in rabbits: a positive cross talk between bradykinin, nitric oxide and prostaglandin E2
Authors: Mello Suzana B. V. , Guzzo Maria Luiza , Lisboa Luiz Filipe Santiago , Farsky Sandra H. P.
Published: November 2003

Background: Our previous results showed that nitric oxide (NO) and bradykinin (BK) mediate the arthritis induced by
Bothrops jararaca venom (BjV) in rabbits. In this study, we investigated the contribution of each receptor of BK as well as the inter‐relationship between NO and eicosanoids in BjV‐induced arthritis.

Methods: The arthritis was induced in rabbits with 16 μg of BjV injected intra‐articularly. Prostaglandin E2
(PGE2), thromboxane B2 (TxB2), leukotriene B4 (LTB4) (radioimmunoassay) and nitrite/nitrate concentrations (NO2/NO3) (Griess reaction) were evaluated in the synovial fluid 4 h later. The animals were prior treated with NO synthase inhibitor (L‐NAME; 20 mg/kg/day for 14 days), the B2 antagonist of BK (HOE‐140) and the B1 antagonist of BK
(des‐Arg9[Leu8]‐bradykinin), both at a dose of 0.3 mg/kg, 30 min prior to the venom injection.

Results: Data show that L‐NAME and HOE‐140 treatment were equally able to reduce PGE2 and NO2/NO3 levels without interfering with TxB2 and LTB4 production. On the contrary, the B1 antagonist of BK inhibited TxB2 and LTB4 production, and did not alter PGE2 and NO metabolites levels in the inflamed joint.

Discussions: The results presented clarify the contribution of the kinin system, mainly through the B2 receptor, to the local inflammatory response induced by BjV, as well as its positive interaction with PGE2 and NO production.
Effect of Tityus serrulatus venom on cytokine production and the activity of murine macrophages
Authors: Petricevich Vera L.
Published: November 2003

The purpose of this study was to investigate the effects of Tityus serrulatus venom (TSV) on murine peritoneal macrophages evaluated in terms of activation. The effects of crude TSV were analysed by detection of cytokines, oxygen intermediate metabolites
(H2O2) and nitric oxide (NO) in supernatants of peritoneal macrophages. Several functional bioassays were employed including an in vitro model for envenomating: cytotoxicity of TSV was assessed using the lyses percentage. Tumor necrosis factor (TNF) activity was assayed by measuring its cytotoxic activity on L‐929 cells, and interleukin‐6 (IL‐6) and interferon‐γ (IFN‐γ) were assayed by enzyme‐linked immunosorbent assay, whereas NO levels were detected by Griess colorimetric reactions in culture supernatant of macrophages incubated with TSV and subsequently exposed to either lipopolysaccharide or IFN‐γ. Incubation of macrophages with TSV increased production of IL‐6 and IFN‐γ in a dose‐dependent manner. TNF production was not detected in supernatants treated with TSV at any concentration. The increase in IL‐6 secretion was not associated with concentration‐dependent cytoxicity of TSV on these cells. These data suggest that the cytotoxicity does not appear to be the main cause of an increased cytokine production by these cells. Although NO is an important effector molecule in macrophage microbicidal activity, the inducing potential of the test compounds for its release was found to be very moderate, ranging from 125 to 800 mM. Interestingly, NO levels of peritoneal macrophages were increased after IFN‐γ. Moreover, NO production had an apparent effect on macrophage activity. The results obtained here also shown that the TSV induces an important elevation in H2O2 release. These results combined with NO production suggest that TSV possesses significant immunomodulatory activities capable of stimulating immune functions in vitro.
Immunomodulating effects of tofizopam (Grandaxin®) and diazepam in vitro
Authors: Kalashnikov Sergey V. , Kalashnikova Elena A. , Kokarovtseva Svetlana N.
Published: November 2003

Benzodiazepines (BDZs) are known to act not only in the central nervous system, but on peripheral cells and tissues binding to the peripheral‐type benzodiazepine receptors. In the present study, the influence of two different BDZs (diazepam (Dz) and tofizopam (Tof)) on several immune functions has been examined in vitro. Some differences between Dz and Tof in their effects on human lymphocyte proliferative response, changes in glucocorticoid‐induced suppression of cell proliferation and influence on cytokine production (tumor necrosis factor‐α (TNF‐α) and interleukin‐2 (IL‐2)) have been determined. Dz suppressed mitogen‐induced peripheral blood mononuclear cell (PBMC) proliferation, enhanced dexamethasone‐induced inhibition of PBMC proliferative response, and suppressed lymphocyte production of TNF‐α and IL‐2. Tof usually enhanced PBMC proliferation and IL‐2 production in low and moderate doses, but in high doses it suppressed both. Tof in all investigated doses enhanced dexamethasone‐induced suppression of lymphocyte proliferation and depressed TNF‐α production. Thus, both Dz and Tof are shown to have immunomodulating effects in vitro. Tof, opposite to Dz even in the therapeutic doses, is able to enhance in vitro mitogen‐induced lymphocyte proliferation and IL‐2 production.
Azelastine and suplatast shorten the distribution half‐life of IgE in rats
Authors: Hanashiro Kazuhiko , Tokeshi Yoshihiro , Nakasone Toshiyuki , Sunagawa Masanori , Nakamura Mariko , Kosugi Tadayoshi
Published: November 2003

We aim to clarify whether suplatast and azelastine (anti‐allergic drugs) can shorten the half‐life of immunoglobulin E (IgE) in the circulating blood. Thirty Wistar rats were divided into six groups. Distilled water or anti‐allergic drugs were given orally for 6 days after the first sensitization. Two milligrams of monoclonal dinitrophenyl (DNP)‐specific rat IgE was administered to the rats, which had been given suplatast or azelastine orally. The level of DNP‐specific rat IgE in the serum was estimated by IgE‐capture enzyme‐linked immunosorbent assay, and the turnover of IgE was analyzed from its pharmacokinetic parameters. The elimination half‐life of rat IgE was about 12 h irrespective of the sensitized state. The intercompartmental rate constants (Kct
and
Ktc) in the suplatast‐administered or azelastine‐administered group were larger than those of the distilled water‐administered group under non‐sensitized conditions. These findings suggested that the anti‐allergic drugs used in the present study facilitated the excretion of IgE from the circulation in rats.
IL‐1 stimulates ceramide accumulation without inducing apoptosis in intestinal epithelial cells
Authors: Homaidan Fadia R. , El-Sabban Marwan E. , Chakroun Iman , El-Sibai Mirvat , Dbaibo Ghassan S.
Published: November 2003

Background: In inflammatory bowel disease (IBD), cytokine levels (such as interleukin‐1 (IL‐1)) are elevated. We have shown previously that IL‐1 activates phospholipid signaling pathways in intestinal epithelial cells (IEC), leading to increased ceramide levels.

Aim: To determine whether ceramide induces apoptosis in IEC.

Methods: Apoptosis was evaluated by annexin‐Vbinding or Hoechst nuclear staining. Levels of bcl‐2, bcl‐x, bax, p53 and p21 were determined by Western blotting, and cell cycle analysis was determined by flow cytometry.

Results: IL‐1 increased ceramide accumulation in a time‐dependent and concentration‐dependent manner with a peak response at 4 h, with [IL‐1] = 30 ng/ml. Neither IL‐1 nor ceramide induced apoptosis in IEC, but they increased bcl‐2 levels and decreased bax and p21 levels without affecting bcl‐x and p53 levels. They also caused a slight but significant increase in the G2/M phase. These data suggest a role for ceramide in IBD and suggest a possible mechanism for the enhanced tumorigenic activity in IBD patients.
High serum level of endostatin in multiple myeloma at diagnosis but not in the plateau phase after treatment
Authors: Urbańska-Ryś Halina , Robak Tadeusz
Published: November 2003

We investigated the serum concentration of endostatin in 84 patients with multiple myeloma (MM) and in 13 healthy controls. The level of measured anti‐angiogenic agent was correlated with the phase and stage of the disease, and most importantly with clinical and laboratory parameters depicting the disease activity (haemoglobin, creatinine, albumins, calcium, M‐component, C‐reactive protein, β2‐microglobulin, lactate dehydrogenase, stage of bone disease) as well as serum levels of pro‐angiogenic cytokines such as vascular endothelial growth factor, hepatocyte growth factor, fibroblast growth factor and transforming growth factor‐β. The median serum level of endostatin in MM patients was 58 ng/ml and was statistically significantly higher than in the control group (median, 40 ng/ml; P = 0.015). MM patients in phase I (at diagnosis) had higher levels of endostatin (median, 69 ng/ml) than those in phase II (plateau phase after treatment) (median, 49 pg/ml; P = 0.044). We did not find any statistical correlation between the level of endostatin and stage of MM according to the Durie and Salmon system. The serum concentration of endostatin in MM patients with a normal level of albumins was significantly higher than in others with hypoalbuminaemia (median, 62 ng/ml versus 39 ng/ml; P = 0.033). Also, patients with a normal value of lactate dehydrogenase had a higher concentration of endostatin than those with values >425 U/l (median, 70 ng/ml versus 39 ng/ml; P = 0.019). We did not show any statistical correlation between the concentration of endostatin and level of haemoglobin, creatinine, calcium, C‐reactive protein, β2‐microglobulin and stage of bone disease. We failed to find positive or negative correlations between the level of endostatin and vascular endothelial growth factor, hepatocyte growth factor, fibroblast growth factor and transforming growth factor‐β. The concentration of endostatin did not influence the probability of survival in MM patients in our study.

In conclusion, our data indicate that endostatin has a higher level in MM patients than in healthy controls. Highest values were stated in active phases of the disease (at presentation and in progression). Different clinical and laboratory parameters generally do not influence the concentration of endostatin (except albumins and lactate dehydrogenase).
Modified guidance laws to escape microbursts with turbulence
Authors: Dogan Atilla , Kabamba Pierre T.
Published: November 2003

This paper introduces Modified Altitude‐ and Dive‐Guidance laws for escaping a microburst with turbulence. The goal is to develop a procedure to estimate the highest altitude at which an aircraft can fly through a microburst without running into stall. First, a new metric is constructed that quantifies the aircraft upward force capability in a microburst encounter. In the absence of turbulence, the metric is shown to be a decreasing function of altitude. This suggests that descending to a low altitude may improve safety in the sense that the aircraft will have more upward force capability to maintain its altitude. In the presence of stochastic turbulence, the metric is treated as a random variable and its probability distribution function is analytically approximated as a function of altitude. This approximation allows us to determine the highest safe altitude at which the aircraft may descend, hence avoiding to descend too low. This highest safe altitude is used as the commanded altitude in Modified Altitude‐ and Dive‐Guidance. Monte Carlo simulations show that these Modified Altitude‐ and Dive‐Guidance strategies can decrease the probability of minimum altitude being lower than a given value without significantly increasing the probability of crash.
Multiperiod hydrothermal economic dispatch by an interior pointmethod
Authors: Kimball L. M. , Clements K. A. , Davis P. W. , Nejdawi I.
Published: November 2003

This paper presents an interior point algorithm to solve the multiperiod hydrothermal economic dispatch (HTED). The multiperiod HTED is a large scale nonlinear programming problem. Various optimization methods have been applied to the multiperiod HTED, but most neglect important network characteristics or require decomposition into thermal and hydro subproblems. The algorithm described here exploits the special bordered block diagonal structure and sparsity of the Newton system for the first order necessary conditions to result in a fast efficient algorithm that can account for all network aspects. Applying this new algorithm challenges a conventional method for the use of available hydro resources known as the peak shaving heuristic.
General solutions of the nonlinear PDEs governing the erosionkinetics
Authors: Panayotounakos D. E. , Zafeiropoulos K. P.
Published: November 2003

We present the construction of the general solutions concerning the one‐dimensional (1D) fully dynamic nonlinear partial differential equations (PDEs), for the erosion kinetics. After an uncoupling procedure of the above mentioned equations a second–order nonlinear PDE of the Monge type governing the porosity is derived, the general solution of which is constructed in the sense that a full complement of arbitrary functions (as many as the order) is introduced. Afterwards, we specify the above solution according to convenient initial conditions.
Global regular solutions for the nonhomogeneous Carrier equation
Authors: Larkin N. A.
Published: November 2003

We study in an + 1‐dimensional cylinder Q global solvability of the mixed problem for the nonhomogeneous Carrier equation

without restrictions on a size of initial data and f(x, t). For any natural n, we prove existence, uniqueness and the exponential decay of the energy for global generalized solutions. When n=2, we prove C∞(Q)‐regularity of solutions.
Boundary value problems on the half line in the theory of colloids
Authors: Agarwal Ravi P. , O′Regan Donal
Published: November 2003

We present existence results for some boundary value problems defined on infinite intervals. In particular our discussion includes a problem which arises in the theory of colloids.
Determination of an unknown parameter in a semi‐linear parabolicequation
Authors: Dehghan Mehdi
Published: November 2003

Some finite difference approximations are developed for an inverse problem of determining an unknown parameter p(t) which is a coefficient of the solution u in a semi‐linear parabolic partial differential equation subject to a boundary integral overspecification. The accuracy and efficiency of the procedures are discussed. Some computational results using the newly proposed numerical techniques are presented. CPU times needed for this problem are reported.
Guided waves in a fluid‐loaded transversely isotropic plate
Authors: Ahmad F. , Kiyani N. , Yousaf F. , Shams M.
Published: November 2003

Dispersion relations are obtained for the propagation of symmetric and antisymmetric modes in a free transversely isotropic plate. Dispersion curves are plotted for the first four symmetric modes for a magnesium plate immersed in water. The first mode is highly damped and switches over to the second mode when the normalized frequency exceeds 12.
FHA Domains as Phospho‐Threonine Binding Modules in Cell Signaling
Authors: Hammet Andrew , Pike Brietta , McNees Carolyn , Conlan Lindus , Tenis Nora , Heierhorst Jörg
Published: November 2003

Forkhead‐associated (FHA) domains are present in <200 diverse proteins in all phyla from bacteria to mammals and seem to be particularly prevalent in proteins with cell cycle control functions. Recent work from several laboratories has considerably improved our understanding of the structure and function of these domains that were virtually unknown a few years ago, and the first disease associations of FHA domains have now emerged. FHA domains form 11‐stranded beta‐sandwiches that contain some 100‐180 amino acid residues with a high degree of sequence diversity. FHA domains act as phosphorylation‐dependent protein‐protein interaction modules that preferentially bind to phospho‐threonine residues in their targets. Interestingly, point mutations in the human CHK2 gene that lead to single‐residue amino acid substitutions in the FHA domain of this cell cycle checkpoint kinase have been found to cause a subset of cases of the Li‐Fraumeni multi‐cancer syndrome. IUBMB Life, 55: 23‐27, 2003
Apoptosis in Yeasts
Authors: Weinberger Martin , Ramachandran Lakshmi , Burhans William
Published: November 2003

Although yeasts lack some elements of the complex apoptotic machinery of metazoan cells, recent studies show that many features of apoptosis, including a caspase‐like activity, can be induced in these organisms by DNA damage and other apoptotic triggers. These remarkable findings provide a compelling argument for increased efforts to bring the powerful genetic approaches available to yeast researchers more directly to bear on questions related to apoptosis and its induction or inhibition by drugs. Yeasts may provide a particularly useful model for understanding connections between DNA damage, cell cycle regulation and apoptosis. Here we summarize these recent findings and explore their implications, particularly for the development of more effective therapeutic strategies for treating cancer. IUBMB Life, 55: 467‐472, 2003
Vigorous Premalignancy-specific Effector T Cell Response in the Bone Marrow of Patients with Monoclonal Gammopathy
Authors: Dhodapkar Madhav V. , Krasovsky Joseph , Osman Keren , Geller Matthew D.
Published: November 2003

Most approaches targeting the immune system against tumors have focused on patients with established tumors. However, whether the immune system can recognize preneoplastic stages of human cancer is not known. Here we show that patients with preneoplastic gammopathy mount a vigorous T cell response to autologous premalignant cells. This preneoplasia-specific CD4+ and CD8+ T cell response is detected in freshly isolated T cells from the BM. T cells from myeloma marrow lack this tumor-specific rapid effector function. These data provide direct evidence for tumor specific immune recognition in human preneoplasia and suggest a possible role for the immune system in influencing the early growth of transformed cells, long before the development of clinical cancer.
Identification of a Pre-BCR Lacking Surrogate Light Chain
Authors: Su Yu-wen , Flemming Alexandra , Wossning Thomas , Hobeika Elias , Reth Michael , Jumaa Hassan
Published: November 2003

SLP-65−/− pre-B cells show a high proliferation rate in vitro. We have shown previously that λ5 expression and consequently a conventional pre-B cell receptor (pre-BCR) are essential for this proliferation. Here, we show that pre-B cells express a novel receptor complex that contains a μ heavy chain (μHC) but lacks any surrogate (SL) or conventional light chain (LC). This SL-deficient pre-BCR (SL−pre-BCR) requires Ig-α for expression on the cell surface. Anti-μ treatment of pre-B cells expressing the SL−pre-BCR induces tyrosine phosphorylation of substrate proteins and a strong calcium (Ca2+) release. Further, the expression of the SL−pre-BCR is associated with a high differentiation rate toward κLC-positive cells. Given that B cell development is only partially blocked and allelic exclusion is unaffected in SL-deficient mice, we propose that the SL−pre-BCR is involved in these processes and therefore shares important functions with the conventional pre-BCR.
Ecsit is required for Bmp signaling and mesoderm formation during mouse embryogenesis
Authors: Xiao Changchun , Shim Jae-hyuck , Klüppel Michael , Zhang Samuel Shao-Min , Dong Chen , Flavell Richard A. , Fu Xin-Yuan , Wrana Jeffrey L. , Hogan Brigid L.M. , Ghosh Sankar
Published: November 2003

Bone morphogenetic proteins (Bmps) are members of the transforming growth factor β (TGFβ) superfamily that play critical roles during mouse embryogenesis. Signaling by Bmp receptors is mediated mainly by Smad proteins. In this study, we show that a targeted null mutation of Ecsit, encoding a signaling intermediate of the Toll pathway, leads to reduced cell proliferation, altered epiblast patterning, impairment of mesoderm formation, and embryonic lethality at embryonic day 7.5 (E7.5), phenotypes that mimic the Bmp receptor type1a (Bmpr1a) null mutant. In addition, specific Bmp target gene expression is abolished in the absence of Ecsit. Biochemical analysis demonstrates that Ecsit associates constitutively with Smad4 and associates with Smad1 in a Bmp-inducible manner. Together with Smad1 and Smad4, Ecsit binds to the promoter of specific Bmp target genes. Finally, knock-down of Ecsit with Ecsit-specific short hairpin RNA inhibits both Bmp and Toll signaling. Therefore, these results show that Ecsit functions as an essential component in two important signal transduction pathways and establishes a novel role for Ecsit as a cofactor for Smad proteins in the Bmp signaling pathway.
tRNAHis maturation: An essential yeast protein catalyzes addition of a guanine nucleotide to the 5′ end of tRNAHis
Authors: Gu Weifeng , Jackman Jane E. , Lohan Amanda J. , Gray Michael W. , Phizicky Eric M.
Published: November 2003

All tRNAHis molecules are unusual in having an extra 5′ GMP residue (G-1) that, in eukaryotes, is added after transcription and RNase P cleavage. Incorporation of this G-1 residue is a rare example of nucleotide addition occurring at an RNA 5′ end in a normal phosphodiester linkage. We show here that the essential Saccharomyces cerevisiae ORF YGR024c (THG1) is responsible for this guanylyltransferase reaction. Thg1p was identified by survey of a genomic collection of yeast GST-ORF fusion proteins for addition of [α-32P]GTP to tRNAHis. End analysis confirms the presence of G-1. Thg1p is required for tRNAHis guanylylation in vivo, because cells depleted of Thg1p lack G-1 in their tRNAHis.His6-Thg1p purified from Escherichia coli catalyzes the guanylyltransferase step of G-1 addition using a ppp-tRNAHis substrate, and appears to catalyze the activation step using p-tRNAHis and ATP. Thg1p is highly conserved in eukaryotes, where G-1 addition is necessary, and is not found in eubacteria, where G-1 is genome-encoded. Thus, Thg1p is the first member of a new family of enzymes that can catalyze phosphodiester bond formation at the 5′ end of RNAs, formally in a 3′-5′ direction. Surprisingly, despite its varied activities, Thg1p contains no recognizable catalytic or functional domains.
Account planning in the new marketing and communications environment (has the Stephen King challenge been met?)
Authors: Zambardino Adrian , Goodfellow John
Published: November 2003

This paper briefly outlines the origins and development of the account planning function within advertising agencies. The transformation of the marketing communications environment during the 1990s is discussed together with the evolution of marketing theory and practice. The main focus of the paper examines if and how account planning has responded to the challenges of the marketing environment during the 1990s and the increasing demand for integrated marketing communications. It questions the extent to which account planning is meeting marketing communications needs in the new millennium both in terms of its traditional internal agency functions and the much debated topic of media neutral planning. The term media neutral planning is analysed and the distinction is drawn between media neutral ideas and neutrality in channel choice. The paper concludes that planning is an inherently flexible discipline and that its prime function is to place the consumer at the heart of creative marketing and communications planning.
Infusing business and brand strategies into account planning
Authors: Osler Rob
Published: November 2003

Account planning has established itself as a core competency of many top advertising agencies. The role of account planner is a result of advertising agencies’ recognition that effective advertising demands the influence of consumer insights, competitive intelligence, and trends analysis. Yet, as necessary as the account planner’s strategic input is to the advertising placement and creative output, it does not supply the prerequisite upstream strategic foundation that is defined and delivered by business and brand strategy. Business strategy must be accounted for to ensure that a firm’s market opportunities, brand opportunities, and core competencies are being fully exploited in all marketing activities. In turn, brand strategy must be leveraged into the account planning process to ensure that a brand is optimally represented within the brand portfolio, its positioning supported, its values reflected, and its personality celebrated in all marketing communications.
Phylogeny of Orbirhynchia Pettitt, 1954 (Brachiopoda: Rhynchonellida)
Authors: Monks Neale , Owen Ellis
Published: November 2003

A computer‐based parsimony analysis of the brachiopod genus Orbirhynchia (Late Albian–Middle Campanian) is described. The resulting cladogram indicates that the genus divided into two distinct lineages soon after its appearance in the Late Albian. One group is known only from marly and sandy facies of Cenomanian age, while the second, more diverse, group appeared later in the Cenomanian but persisted into the Campanian. These two groups exhibit distinct morphological trends which may be related to different ecological niches, perhaps with respect to local sedimentary environment. Key words: brachiopod, Orbirhynchia, Cretaceous, phylogeny, palaeoecology.
Permineralized Cardiocarpalean Ovules in Wetland Vegetation from Early Permian Volcaniclastic Sediments of China
Authors: Hilton Jason , Rothwell Gar W. , Li Cheng‐sen , Wang Shi‐jun , Galtier Jean
Published: November 2003

In this paper we report the first occurrence of permineralized plant fossils in volcaniclastic lithologies from China and in doing so describe a new species of cardiocarpalean ovule permineralized within tuffaceous sediments from a recently discovered locality near Pingquan, Hebei Province, North China. The tuff is part of the Early Permian Taiyuan Formation which contains a diverse fossil plant assemblage that includes cordaitean spermatophytes, lepidodendralean lycophytes, equisetophytes, and filicalean ferns, all of which were typical of Permian floras of northern China at this time. Specimens of Cardiocarpusdabiziae sp. nov. have the characteristic platyspermic shape and vascularization of cardiocarpalean ovules, and display prominent protuberances on the exterior of the integument. The integument of the ovule is composed of three layers; a thick sarcotesta, a thin and comparatively dense sclerotesta, and a single layer of large endotesta cells. The pollen chamber produces a slender nucellar beak. In several specimens the megaspore membrane contains well‐developed tissue of the megagametophyte. The ovules have prominent external integumentary protuberances which suggest that well‐developed systems of plant/animal interactions were operative in Early Permian wetland biotas of the Cathaysian realm.
Activation of bronchopulmonary vagal afferent nerves with bradykinin, acid and vanilloid receptor agonists in wild‐type and TRPV1–/– mice
Authors: Kollarik M. , Undem B. J.
Published: November 2003

The vanilloid receptor TRPV1 (formerly VR1) has been implicated in the activation of nociceptive sensory nerves by capsaicin, noxious heat, protons, bradykinin, cannabinoids such as anandamide, and certain metabolites of arachidonic acid. Using TRPV1 knockout mouse (TRPV1–/–) we address the question of whether TRPV1 is obligatory for action potential discharge in vagal C‐fibre terminals evoked by capsaicin, anandamide, acid and bradykinin. The response of a defined subtype of the vagal afferent bronchopulmonary C‐fibres (conduction velocity < 0.7 ms−1) to the putative TRPV1 activators was studied in vitro in the mouse isolated/perfused lung–nerve preparation. Capsaicin (1 μm) evoked action potential discharge of ∼90% (28/31) of C‐fibres in the TRPV1+/+ mice, but failed to activate bronchopulmonary C‐fibres in TRPV1–/– animals (n= 10). Anandamide (3–100 μm) induced concentration‐dependent activation of capsaicin‐sensitive TRPV1+/+ C‐fibres with a threshold of 3–10 μm, but failed to evoke substantive discharge in TRPV1–/– C‐fibres. In the TRPV1+/+ mice, the B2 receptor‐mediated activation by bradykinin (1 μm) was restricted to the capsaicin‐sensitive C‐fibres. Bradykinin was effective in evoking B2 receptor‐mediated action potential discharge in TRPV1–/– C‐fibres, but the response was significantly (P < 0.05) less persistent than in TRPV1+/+ C‐fibres. Exposing the tissue to acid (pH = 5) excited both TRPV1+/+ and TRPV1–/– C‐fibres. We conclude that TRPV1 is obligatory for vagal C‐fibre activation by capsaicin and anandamide. By contrast, whereas TRPV1 may have a modulatory role in bradykinin and acid‐induced activation of bronchopulmonary C‐fibres, it is not required for action potential discharge evoked by these stimuli.
Effects of acute and chronic endurance exercise on mitochondrial uncoupling in human skeletal muscle
Authors: Fernström Maria , Tonkonogi Michail , Sahlin Kent
Published: November 2003

Mitochondrial proteins such as uncoupling protein 3 (UCP3) and adenine nucleotide translocase (ANT) may mediate back‐leakage of protons and serve as uncouplers of oxidative phosphorylation. We hypothesized that UCP3 and ANT increase after prolonged exercise and/or endurance training, resulting in increased uncoupled respiration (UCR). Subjects were investigated with muscle biopsies before and after acute exercise (75 min of cycling at 70% of ) or 6 weeks endurance training. Mitochondria were isolated and respiration measured in the absence (UCR or state 4) and presence of ADP (coupled respiration or state 3). Protein expression of UCP3 and ANT was measured with Western blotting. After endurance training , citrate synthase activity (CS), state 3 respiration and ANT increased by 24, 47, 40 and 95%, respectively (all P < 0.05), whereas UCP3 remained unchanged. When expressed per unit of CS (a marker of mitochondrial volume) UCP3 and UCR decreased by 54% and 18%(P < 0.05). CS increased by 43% after acute exercise and remained elevated after 3 h of recovery (P < 0.05), whereas the other muscle parameters remained unchanged. An intriguing finding was that acute exercise reversibly enhanced the capacity of mitochondria to accumulate Ca2+(P < 0.05) before opening of permeability transition pores. In conclusion, UCP3 protein and UCR decrease after endurance training when related to mitochondrial volume. These changes may prevent excessive basal thermogenesis. Acute exercise enhances mitochondrial resistance to Ca2+ overload but does not influence UCR or protein expression of UCP3 and ANT. The increased Ca2+ resistance may prevent mitochondrial degradation and the mechanism needs to be further explored.
Nuclear Export of MicroRNA Precursors
Authors: Lund Elsebet , Güttinger Stephan , Calado Angelo , Dahlberg James E. , Kutay Ulrike
Published: November 2003

MicroRNAs (miRNAs), which function as regulators of gene expression in eukaryotes, are processed from larger transcripts by sequential action of nuclear and cytoplasmic ribonuclease III–like endonucleases. We show that Exportin-5 (Exp5) mediates efficient nuclear export of short miRNA precursors (pre-miRNAs) and that its depletion by RNA interference results in reduced miRNA levels. Exp5 binds correctly processed pre-miRNAs directly and specifically, in a Ran guanosine triphosphate–dependent manner, but interacts only weakly with extended pre-miRNAs that yield incorrect miRNAs when processed by Dicer in vitro. Thus, Exp5 is key to miRNA biogenesis and may help coordinate nuclear and cytoplasmic processing steps.
High-Performance Solid Acid Fuel Cells Through Humidity Stabilization
Authors: Boysen Dane A. , Uda Tetsuya , Chisholm Calum R. I. , Haile Sossina M.
Published: November 2003

Although they hold the promise of clean energy, state-of-the-art fuel cells based on polymer electrolyte membrane fuel cells are inoperable above 100°C, require cumbersome humidification systems, and suffer from fuel permeation. These difficulties all arise from the hydrated nature of the electrolyte. In contrast, “solid acids” exhibit anhydrous proton transport and high-temperature stability. We demonstrate continuous, stable power generation for both H
2
/O
2
and direct methanol fuel cells operated at ∼250°C using a humidity-stabilized solid acid CsH
2
PO
4
electrolyte.
Genetic Basis for Biosynthesis of the(α1→4)-Linked
N
-Acetyl-
d
-Glucosamine 1-Phosphate Capsuleof
Neisseria meningitidis
SerogroupX
Authors: Tzeng Yih-Ling , Noble Corie , Stephens David S.
Published: November 2003

The
genetic basis for biosynthesis of the (α1→4)-linked
N
-acetyl-
d
-glucosamine 1-phosphate capsule of
Neisseria meningitidis
serogroup X was defined. The
biosynthesis gene cassette was a ∼4.2-kb region located between
ctrA
of the capsule transport operon and
galE
, which
encodes the UDP-glucose-4-epimerase. This location was identical to the
locations of the biosynthesis cassettes in other meningococcal
serogroups. Three open reading frames unique to meningococcus serogroup
X were identified. Deletion-insertion mutation and colony
immunoblotting confirmed that these three genes were essential for
serogroup X capsule expression, and the genes were designated
xcbA
,
xcbB
, and
xcbC
(serogroup
X
c
apsule
b
iosynthesis). Reverse transcriptase PCR
indicated that the
xcbABC
genes form an operon and are
cotranscribed divergently from
ctrA
. XcbA exhibited 52%
amino acid similarity to SacB, the putative capsule polymerase of
meningococcus serogroup A, suggesting that it plays a role as the
serogroup X capsule polymerase. An IS
1016
element was found
within the intergenic region separating
ctrA
and
xcbA
in multiple strains, and this element did not interfere with capsule
expression.
Association of
Salmonella enterica
SerovarEnteritidis YafD with Resistance to Chicken EggAlbumen
Authors: Lu Sangwei , Killoran Patrick B. , Riley Lee W.
Published: November 2003

Salmonella
enterica
serovar Enteritidis is a major cause of food-borne
diseases in industrialized countries. The incidence of
S.
enterica
serovar Enteritidis infections has increased
substantially in recent decades, and
S. enterica
serovar
Enteritidis is now one of the leading serovars of
Salmonella
in the United States. A unique epidemiological characteristic of
S.
enterica
serovar Enteritidis is its association with chicken shell
eggs, since approximately 80% of all human gastrointestinal
diseases can be traced to contaminated egg products. Eggs are
contaminated when bacteria from reproductive tissues of infected hens
are packaged into the eggs and persist inside the hostile egg albumen
environment. Therefore, resistance to egg albumen is an important
aspect in the transmission of
S. enterica
serovar Enteritidis.
We identified a gene,
yafD
from
S. enterica
serovar
Enteritidis, whose overexpression conferred upon
S. enterica
serovar Typhimurium enhanced resistance to egg albumen, while
disruption of this gene in
S. enterica
serovar Enteritidis
rendered the organism more susceptible to egg albumen. YafD is
homologous to members of an exonuclease-endonuclease-phosphatase
family, including some enzymes involved in DNA repair. Furthermore, we
discovered that egg albumen has nuclease activities and uses both
circular and linear DNA as substrates. We propose that YafD provides a
survival advantage to
S. enterica
serovar Enteritidis in eggs
by repairing DNA damage caused by egg albumen and that it may be one of
the biologic determinants that contribute to the epidemiological
association of
S. enterica
serovar Enteritidis with egg
products.
Lipopolysaccharide Binding Protein Is an Essential Component of the Innate Immune Response toEscherichia coliPeritonitis in Mice
Authors: Knapp Sylvia , de Vos Alex F. , Florquin Sandrine , Golenbock Douglas T. , van der Poll Tom
Published: November 2003

Lipopolysaccharide (LPS) binding protein (LBP) is an acute-phase protein that enhances the responsiveness of immune cells to LPS by virtue of its capacity to transfer LPS to CD14. To determine the role of LBP in the innate immune response to peritonitis, LBP gene-deficient (LBP−/−) and normal wild-type mice were intraperitoneally infected withEscherichia coli, the most common causative pathogen of this disease. LBP was detected at low concentrations in peritoneal fluid of healthy wild-type mice, and the local LBP levels increased rapidly upon induction of peritonitis. LBP−/−mice were highly susceptible toE. coliperitonitis, as indicated by accelerated mortality, earlier bacterial dissemination to the blood, impaired bacterial clearance in the peritoneal cavity, and more severe remote organ damage. LBP−/−mice displayed diminished early tumor necrosis factor alpha, interleukin-6, cytokine-induced neutrophil chemoattractant, and macrophage inflammatory protein 2 production and attenuated recruitment of polymorphonuclear leukocytes to the site of infection, indicating that acute inflammation was promoted by LBP. Locally produced LBP is an essential component of an effective innate immune response toE. coliperitonitis.
Characterization and Development of T-Cell Immune Responses in B-Cell-Deficient (Igh-6−/−) Mice withSalmonella entericaSerovar TyphimuriumInfection
Authors: Ugrinovic Sanja , Ménager Nathalie , Goh Natalie , Mastroeni Pietro
Published: November 2003

Infection of mice withSalmonella entericaserovar Typhimurium induces strong Th1 T-cell responses that are central to the control of the infection. In the present study, we examined the role of B cells in the development of Th1 T-cell responses toSalmonellaby using gene-targeted B-cell-deficient mice (Igh-6−/−mice). The development of Th1 T-cell responses inIgh-6−/−mice was impaired in the early stage of a primary infection. This impairment persisted throughout the course of the disease. The ability of T cells to produce the Th1 cytokine gamma interferon and the frequency at which they did so were lower inIgh-6−/−mice than in control mice. We also observed a transient switch toward Th2 cytokine production inIgh-6−/−mice. Thus, B cells are important for the induction of protective Th1 T-cell responses in the early phase of aSalmonellainfection. Activated B cells express high levels of major histocompatibility complex and costimulatory molecules and are nearly as effective as dendritic cells in their antigen-presenting cell (APC) activity. However, their importance as APCs in infection and their role in initiating and/or maintaining T-cell responses are unknown. Here, we show that B cells upregulate costimulatory molecules upon in vitro stimulation withS. entericaserovar Typhimurium and that they can presentSalmonellaantigens toSalmonella-specific CD4+T cells. Our results show that B cells are important for the development of T-cell responses in the early stage of aSalmonellainfection and that this property may be due to their ability to present antigens to T cells.
Resistance and Susceptibility to Filarial InfectionwithLitomosoides sigmodontisAre Associated with EarlyDifferences in Parasite Development and in Localized ImmuneReactions
Authors: Babayan Simon , Ungeheuer Marie-Noëlle , Martin Coralie , Attout Tarik , Belnoue Elodie , Snounou Georges , Rénia Laurent , Korenaga Masataka , Bain Odile
Published: November 2003

In order to understand natural resistance to filariasis, we comparedLitomosoides sigmodontisprimary infection of C57BL/6 mice, which eliminate the worms before patency, and BALB/c mice, in which worms complete their development and produce microfilariae. Our analysis over the first month of infection monitoredmigration of the infective larvae from the lymph nodes to the pleural cavity, where the worms settle. Although immune responses from the mouse strains differed from the outset, the duration of lymphatic migration (4 days) and filarial recovery rates were similar, thus confirming that the proportion of larvae that develop in the host species upon infection is not influenced by host genetic variability. The majority of worms reached the adult stage in both mouse strains; however, worm growth and molting were retarded in resistant C57BL/6 mice. Surprisingly, the only immune responses detected at 60 h postinfection occurred in the susceptible mice and only upon stimulation of cells from lymph nodes draining the inoculation site with infective larva extract: massive production of interleukin-6 (IL-6) and IL-5 (the latter cytokine was previously suspected to have an effect onL. sigmodontisgrowth). However, between days 10 and 30 postinfection, extraordinarily high levels of type 1 and type 2 cytokines and expansion of pleural leukocyte infiltration were seen in the resistant C57BL/6 mice, explaining the destruction of worms later. Our results suggest that events early in the infection determine susceptibility or resistance to subsequent microfilarial production and a parasite strategy to use specific immune responses to its own benefit.
The Site ofLeishmania majorInfection Determines Disease Severity andImmuneResponses
Authors: Baldwin Tracey M. , Elso Colleen , Curtis Joan , Buckingham Lynn , Handman Emanuela
Published: November 2003

Inbred strains of mice infected withLeishmania majorhave been classified as genetically resistant or susceptible on the basis of their ability to cure their lesions, the parasite burden in the draining lymph nodes, and their type of T helper cell immune responses to the parasite. Using the intradermal infection at the base of the tail and the ear pinna, we compared for the first time the above-mentioned parameters in six strains of mice infected with metacyclic promastigotes, and we show that the severity of disease depends greatly on the site of infection. Although the well-documented pattern of disease susceptibility of BALB/c and C57BL/6 mice described for the footpad and base-of-the-tail models of leishmaniasis were confirmed, C3H/HeN and DBA/2 mice, which are intermediate and susceptible, respectively, in the tail and other models, were resistant to ear infection. Moreover, in the CBA/H, C3H/HeN, C57BL/6J, and DBA/2 mouse strains, there was little correlation between the pattern of cytokines produced and the disease phenotype observed at the ear and tail sites. We conclude that the definition of susceptibility and the immune mechanisms leading to susceptibility or resistance to infection may differ substantially depending on the route of infection.