bluuebunny/crossref_metadata_2025 huggingface dataset:
Where poetry points: deixis and poetry's ‘you’ in Éluard and Desnos
Authors: Hochman Hugh
Published: June 2005

The struggle in poetics between the textual hermeticism of semiotic readings, on the one hand, and materialist theories of reference, on the other, has been a thorny issue for theorists of the lyric for some time. Taking up the question of whether or not poetic texts open on to a world of referents, this article considers deictics (words for showing or indicating) in poems by Paul Éluard and Robert Desnos, recalling Émile Benveniste's inclusion of the personal pronouns
I
and
you
, along with demonstratives, under the rubric of
indicateurs.
Specifically, the article examines the function of words such as
this, that
and
you
and looks into the possibility that poems are always addressed to their reader and therefore mean
you
in the strongest sense possible. Poems do not refer to the world mentioned on the level of their themes or fiction but rather perpetually refer to relationships they sustain with the reader. In this sense, the reader's understanding or interpretation is an extension of a poem's addressivity, a kind of intimacy whereby the reader manages to meet the requirements of love and desire according to a uniquely discursive reality. It is precisely in understanding how a poem's semiotic operations carry within them their own justification that the reader fleshes out the addressee or the
you
of the text.
Full Lagrangian methods for calculating particle concentration fields in dilute gas-particle flows
Authors: Healy D.P , Young J.B
Published: June 2005

The paper discusses two
Full Lagrangian
methods for calculating the particle concentration and velocity fields in dilute gas-particle flows. The methods are mathematically similar but crucially different in application. By examining the analytical solution for two-phase stagnation-point flow, it is shown that Osiptsov's method is more general than that of Fernandez de la Mora and Rosner. In Osiptsov's method, the Jacobian of the Eulerian–Lagrangian transformation is computed by integration along particle pathlines. The particle concentration is then obtained algebraically from the Lagrangian form of the particle continuity equation. It is shown that the correct specification of the initial conditions is non-trivial and of vital importance. A technique to alleviate problems of mathematical ‘stiffness’ at small Stokes numbers is also described. Full Lagrangian methods require knowledge of the fluid velocity gradient field and, if the carrier flowfield is calculated numerically, differentiation of a ‘noisy’ field can result in serious errors. The paper describes a method for reducing these errors. The incompressible, inviscid flow over a cylinder provides a useful test case for validation and the Osiptsov method proves its worth by revealing a region, hitherto unknown, of crossing particle pathlines in the mathematical solution. Crossing pathlines and their relationship to Robinson's integral are then discussed, and calculations of particle flow through a turbine cascade at high Mach numbers are presented to illustrate the engineering potential of the method.
Stokes-multiplier expansion in an inhomogeneous differential equation with a small parameter
Authors: Ólafsdóttir E.I , Olde Daalhuis A.B , Vanneste J
Published: June 2005

Accurate approximations to the solutions of a second-order inhomogeneous equation with a small parameter
ϵ
are derived using exponential asymptotics. The subdominant homogeneous solutions that are switched on by an inhomogeneous solution through a Stokes phenomenon are computed. The computation relies on a resurgence relation, and it provides the
ϵ
-dependent Stokes multiplier in the form of a power series. The
ϵ
-dependence of the Stokes multiplier is related to constants of integration that can be chosen arbitrarily in the WKB-type construction of the homogeneous solution.

The equation under study governs the evolution of special solutions of the Boussinesq equations for rapidly rotating, strongly stratified fluids. In this context, the switching on of subdominant homogeneous solutions is interpreted as the generation of exponentially small inertia–gravity waves.
Unparallel diversification in bacterial microcosms
Authors: Tyerman Jabus , Havard Naomi , Saxer Gerda , Travisano Michael , Doebeli Michael
Published: June 2005

Adaptive speciation has gained popularity as a fundamental process underlying the generation of diversity. We tested whether populations respond to similar forms of disruptive selection by diversifying in similar or parallel ways by investigating diversified populations of
Escherichia coli
B evolved in glucose and glucose–acetate environments. In both environments, the populations have differentiated into two phenotypes, named for their characteristic colony morphologies: large (L) and small (S). Each type is heritable and this polymorphism (or ‘diversified pair’) appears to be maintained by negative frequency dependence. The L and S phenotypes from different environments are convergent in their colony morphology and growth characteristics. We tested whether diversification was parallel by conducting competition experiments between L and S types from different environments. Our results indicate that replicate diversified pairs from different environments have not diversified in parallel ways and suggest that subtle differences in evolutionary environment can crucially affect the outcome of adaptive diversification.
Evidence for pollinator sharing in Mediterranean nectar-mimic orchids: absence of premating barriers?
Authors: Cozzolino Salvatore , Schiestl Florian P , Müller Andreas , De Castro Olga , Nardella Antonio Marco , Widmer Alex
Published: June 2005

Pollinator specificity has traditionally been considered the main reproductive isolation mechanism in orchids. Among Mediterranean orchids, however, many species attract and deceive pollinators by mimicking nectar-rewarding plants. To test the extent to which deceptive orchid species share pollinators, we collected and identified hemipollinaria-carrying insects, and used ribosomal sequences to identify the orchid species from which hemipollinaria were removed. We found that social and solitary bees, and also flies, carried hemipollinaria belonging to nine orchid species with different degrees of specialization. In particular,
Anacamptis morio
,
Dactylorhiza romana
and
Orchis mascula
used a large set of pollinator species, whereas others such as
Orchis quadripunctata
seemed to be pollinated by one pollinator species only. Out of the insects with hemipollinaria, 19% were found to carry hemipollinaria from more than one orchid species, indicating that sympatric food-deceptive orchids can share pollinators. This sharing was apparent even among orchid sister-species, thus revealing an effective overlap in pollinator sets among closely related species. These results suggest varying degrees of pollinator specificity in these orchids, and indicate that pollinator specificity cannot always act as the main isolation mechanism in food-deceptive temperate orchids.
Genomic compatibility occurs over a wide range of parental genetic similarity in an outcrossing plant
Authors: Willi Yvonne , Van Buskirk Josh
Published: June 2005

The theory of inbreeding and outbreeding suggests that there is a hump-shaped relationship between the genetic similarity of sexually reproducing parents and the performance of their offspring. Inbreeding depression occurs when genetic similarity is high, whereas hybrid breakdown is expected when genetic similarity is low. Between these extremes, the effect of genetic similarity on fitness is unclear. We studied the shape of this relationship by crossing 65 target genotypes of the clonal, self-incompatibleRanunculus reptanswith partner genotypes spanning a broad scale of genetic similarity, ranging from crosses within populations to between-population crosses and hybridisation with a closely related species. Offspring were raised in outdoor tubs. Results revealed a quadratic relationship between parental genetic distance and offspring performance, with the clonal component of fitness more strongly hump-shaped than the sexual component. Optimal genetic similarity encompassed a broad range of within-population and between-population crosses. This pattern of genomic compatibility has important implications for the evolution of mating systems and mate choice.
Modelling waves and surges during the 1953 storm
Authors: Wolf J , Flather R.A
Published: June 2005

Waves and sea levels have been modelled for the storm of 31 January–1 February 1953. Problems in modelling this event are associated with the difficulty of reconstructing wind fields and validating the model results with the limited data available from 50 years ago. The reconstruction of appropriate wind fields for surge and wave models is examined. The surges and waves are reproduced reasonably well on the basis of tide-gauge observations and the sparse observational information on wave heights. The maximum surge coincided closely in time with tidal high water, producing very high water levels along the coasts of the southern North Sea. The statistics of the 1953 event and the likelihood of recurrence are also discussed. Both surge and wave components were estimated to be approximately 1 in 50 year events. The maximum water level also occurred when the offshore waves were close to their maximum. The estimation of return period for the total water level is more problematic and is dependent on location. A scenario with the 1953 storm occurring in 2075, accounting for the effects of sea level rise and land movements, is also constructed, suggesting that sea level relative to the land could be 0.4–0.5 m higher than in 1953 in the southern North Sea, assuming a rise in mean sea level of 0.4 m.
Full-genome analysis of a highly divergent simian T-cell lymphotropic virus type 1 strain in Macaca arctoides
Authors: Van Dooren Sonia , Meertens Laurent , Lemey Philippe , Gessain Antoine , Vandamme Anne-Mieke
Published: June 2005

Full-genome sequencing and analysis of the highly divergent simian T-cell lymphotropic virus type 1 (STLV-1) strain MarB43 inMacaca arctoidesindicated that its open reading frame structure is compatible with proper functioning of its Gag, Pol, Env, Tax and Rex proteins. Detailed analysis of the coding potential, however, revealed that MarB43 is probably forced to use the human T-cell lymphotropic virus type 2/STLV-2env-tax-rexsplice-acceptor homologue and that the proximal pX auxiliary proteins p12I, p13II, p30IIand p27Iseem to have lost their function. Full-genome (gag-pol-env-tax), long terminal repeat andenvphylogenetic analyses conclusively identified STLV-1 inM. arctoidesas the currently most divergent STLV-1 strain. The long branching pattern of the monophyletic STLV-1Macacasubspecies clades suggests that macaques might be the ancestral reservoir for primate T-cell lymphotropic virus type 1 in Asia. Full-genome molecular-clock analysis supports an archaic introduction of STLV-1 on the Asian continent, at least 269 000–156 000 years ago.
Interaction between the respiratory syncytial virus G glycoprotein cytoplasmic domain and the matrix protein
Authors: Ghildyal Reena , Li Dongsheng , Peroulis Irene , Shields Benjamin , Bardin Phillip G. , Teng Michael N. , Collins Peter L. , Meanger Jayesh , Mills John
Published: June 2005

Paramyxovirus assembly at the cell membrane requires the movement of viral components to budding sites and envelopment of nucleocapsids by cellular membranes containing viral glycoproteins, facilitated by interactions with the matrix protein. The specific protein interactions during assembly of respiratory syncytial virus (RSV) are unknown. Here, the postulated interaction between the RSV matrix protein (M) and G glycoprotein (G) was investigated. Partial co-localization of M with G was demonstrated, but not with a truncated variant lacking the cytoplasmic domain and one-third of the transmembrane domain, in cells infected with recombinant RSV or transfected to express G and M. A series of G mutants was constructed with progressively truncated or modified cytoplasmic domains. Data from co-expression in cells and a cell-free binding assay showed that the N-terminal aa 2–6 of G play a key role in G–M interaction, with serine at position 2 and aspartate at position 6 playing key roles.
New sequence polymorphisms in the outer loops of the JC polyomavirus major capsid protein (VP1) possibly associated with progressive multifocal leukoencephalopathy
Authors: Zheng Huai-Ying , Takasaka Tomokazu , Noda Kazuyuki , Kanazawa Akira , Mori Hideo , Kabuki Tomoyuki , Joh Kohsuke , Oh-ishi Tsutomu , Ikegaya Hiroshi , Nagashima Kazuo , Hall William W. , Kitamura Tadaichi , Yogo Yoshiaki
Published: June 2005

JC polyomavirus(JCPyV) causes progressive multifocal leukoencephalopathy (PML) in patients with decreased immune competence. To elucidate genetic changes in JCPyV associated with the pathogenesis of PML, multiple complete JCPyV DNA clones originating from the brains of three PML cases were established and sequenced. Although unique rearranged control regions occurred in all clones, a low level of nucleotide variation was also found in the coding region. In each case, a parental coding sequence was identified, from which variant coding sequences with nucleotide substitutions would have been generated. A comparison between the parental and variant coding sequences demonstrated that all 12 detected nucleotide substitutions gave rise to amino acid changes. Interestingly, seven of these changes were located in the surface loops of the major capsid protein (VP1). Finally, 16 reported VP1 sequences of PML-type JCPyV (i.e. derived from the brain or cerebrospinal fluid of PML patients) were compared with their genotypic prototypes, generated as consensus sequences of representative archetypal isolates belonging to the same genotypes; 13 VP1 proteins had amino acid changes in the surface loops. In contrast, VP1 proteins from isolates from the urine of immunocompetent and immunosuppressed patients rarely underwent mutations in the VP1 loops. The present findings suggest that PML-type JCPyV frequently undergoes amino acid substitutions in the VP1 loops. These polymorphisms should serve as a new marker for the identification of JCPyV isolates associated with PML. The biological significance of these mutations, however, remains unclear.
Evolutionary dynamics of hepatitis C virus envelope genes during chronic infection
Authors: Brown Richard J. P. , Juttla Vicky S. , Tarr Alexander W. , Finnis Rebecca , Irving William L. , Hemsley Shelley , Flower Darren R. , Borrow Persephone , Ball Jonathan K.
Published: June 2005

Hepatitis C virus (HCV) envelope glycoproteins E1 and E2 are important targets for the host immune response. The genes encoding these proteins exhibit a high degree of variability that gives rise to differing phenotypic traits, including alterations in receptor-binding affinity and immune recognition and escape. In order to elucidate patterns of adaptive evolution during chronic infection, a panel of full-length E1E2 clones was generated from sequential serum samples obtained from four chronically infected individuals. By using likelihood-based methods for phylogenetic inference, the evolutionary dynamics of circulating HCV quasispecies populations were assessed and a site-by-site analysis of thedN/dSratio was performed, to identify specific codons undergoing diversifying positive selection. HCV phylogenies, coupled with the number and distribution of selected sites, differed markedly between patients, highlighting that HCV evolution during chronic infection is a patient-specific phenomenon. This analysis shows that purifying selection is the major force acting on HCV populations in chronic infection. Whilst no significant evidence for positive selection was observed in E1, a number of sites under positive selection were identified within the ectodomain of the E2 protein. All of these sites were located in regions hypothesized to be exposed to the selective environment of the host, including a number of functionally defined domains that have been reported to be involved in immune evasion and receptor binding. Dated-tip methods for estimation of underlying HCV mutation rates were also applied to the data, enabling prediction of the most recent common ancestor for each patient's quasispecies.
Phage display-selected single-chain antibodies confer high levels of resistance against Tomato spotted wilt virus
Authors: Prins Marcel , Lohuis Dick , Schots Arjen , Goldbach Rob
Published: June 2005

Rational design of antibodies targeting essential viral proteins can complement the palette of antiviral resistance strategies. Here, stable and high expression of single-chain monoclonal antibodies targeting the nucleoprotein of the economically important plant virus Tomato spotted wilt virus, a protein that is involved in multiple steps in the viral infection cycle, is reported. High cytoplasmic expression levels of three selected phage display-derived anti-viral single-chain antibodies were established. Of these antibodies, two led to high levels of resistance against this plant virus. Protoplast experiments provided evidence that the two resistance-conferring antibodies may have a different mode of action and could be combined for higher durability of resistance in the field.
Immune responses of pigs after experimental infection with a European strain of Porcine reproductive and respiratory syndrome virus
Authors: Díaz I. , Darwich L. , Pappaterra G. , Pujols J. , Mateu E.
Published: June 2005

The purpose of this experiment was to study the immune response of pigs during an experimental infection with a European strain ofPorcine reproductive and respiratory syndrome virus(PRRSV). Five pigs were challenged intranasally with PRRSV strain VP21 and another five were kept as controls. Clinical course and humoral and cell-mediated responses were monitored for 70 days post-infection (p.i.). Infected pigs developed mild signs at 24 h p.i. Viraemia was detectable by nested RT-PCR until day 14 p.i. Earliest seroconversions (ELISA) were seen by day 7 p.i. (three of five animals) and, by day 14, all inoculated pigs had seroconverted (ELISA and immunoperoxidase monolayer assay). Virus-neutralizing antibodies were undetectable until day 56 p.i. and, by day 70 p.i., two inoculated pigs still were negative. Flow-cytometry assays using peripheral blood mononuclear cells (PBMC) showed an upshift in CD8+cells (day 7 p.i.) and a downshift of CD21+cells (days 7 and 28 p.i.). Regarding cell-mediated responses, development of PRRSV-specific gamma interferon-secreting cells (IFN-γ-SC) and interleukin 4-secreting cells (IL4-SC) in PBMC was examined by ELISPOT assay. IFN-γ-SC were not detected significantly until day 14 p.i., whereas, for IL4-SC, no differences between groups were seen. Concurrently with the onset of viraemia and the development of clinical signs, serum haptoglobin levels and interleukin 10 (IL10) in PRRSV-stimulated PBMC-culture supernatants increased significantly. These differences disappeared later on. For IL2, IL4, IL8 or transforming growth factor beta, no differences were seen among groups. These results are compatible with a model in which the immune response does not fully control the outcome of the infection.
Paramyxovirus mRNA editing, the ‘rule of six’ and error catastrophe: a hypothesis
Authors: Kolakofsky Daniel , Roux Laurent , Garcin Dominique , Ruigrok Rob W. H.
Published: June 2005

The order Mononegavirales includes three virus families that replicate in the cytoplasm: the Paramyxoviridae, composed of two subfamilies, the Paramyxovirinae and Pneumovirinae, the Rhabdoviridae and the Filoviridae. These viruses, also called non-segmented negative-strand RNA viruses (NNV), contain five to ten tandemly linked genes, which are separated by conserved junctional sequences that act as mRNA start and poly(A)/stop sites. For the NNV, downstream mRNA synthesis depends on termination of the upstream mRNA, and all NNV RNA-dependent RNA polymerases reiteratively copy (‘stutter’ on) a short run of template uridylates during transcription to polyadenylate and terminate their mRNAs. The RNA-dependent RNA polymerase of a subset of the NNV, all members of the Paramyxovirinae, also stutter in a very controlled fashion to edit their phosphoprotein gene mRNA, and Ebola virus, a filovirus, carries out a related process on its glycoprotein mRNA. Remarkably, all viruses that edit their phosphoprotein mRNA are also governed by the ‘rule of six’, i.e. their genomes must be of polyhexameric length (6n+0) to replicate efficiently. Why these two seemingly unrelated processes are so tightly linked in the Paramyxovirinae has been an enigma. This paper will review what is presently known about these two processes that are unique to viruses of this subfamily, and will discuss whether this enigmatic linkage could be due to the phenomenon of RNA virus error catastrophe.
Musca domestica is not a vector of Thelazia callipaeda in experimental or natural conditions
Authors: Otranto D. , Lia R. P. , Testini G. , Milillo P. , Shen J. L. , Wang Z. X.
Published: June 2005

Abstract.  Thelazia callipaeda Railliet and Henry (Spirurida: Thelaziidae), commonly called oriental eyeworm for its widespread presence in the Far East, has been recently found to affect dogs, cats and foxes in northern and southern Italy. Although the biology of T. callipaeda in the definitive hosts has been recently investigated, many doubts still remain about its biology in insect vectors. It has been suggested that more than one species of Diptera, namely Musca domestica Linnaeus (Diptera: Muscidae) and Amiota okadai Maca (Diptera: Drosophilidae), may be involved in the transmission of T. callipaeda in China. The aim of the work described here was to verify the role of M. domestica as a vector of T. callipaeda both in experimental and natural conditions. A total of 310 m. domestica (Group 1) were put in a cage and allowed to feed for 14 days around the eyes of a dog naturally infected by T. callipaeda. Ten flies were collected daily for 14 days. A total of 149 houseflies (Group 2) were fed with T. callipaeda first stage larvae (L1) and dissected at 1, 2 and 7 days post‐infection. From June to August 2003, flies were netted (Group 3) in two different sites every 10 days both from the environment and directly from the periocular region of dogs affected by thelaziosis. Musca flies were examined for eyeworms by dissection and visual inspection of house flies (Groups 1 and 2) and using a molecular approach (Groups 1–3) via a specific amplification of the ribosomal internal transcribed spacer 1 (ITS1) sequence of T. callipaeda. On the whole, 180 pools of M. domestica flies were processed molecularly and all the experimentally infected flies (Groups 1 and 2) were found to be negative both at the visual dissection and at the molecular assay. Similarly, the 234 M. domestica collected from Group 3 were negative for T. callipaeda. The results clearly suggest that M. domestica is unlikely to act as a vector of T. callipaeda in southern Europe, in contrast with a single previous report.
A comparison of the susceptibility of Culicoides imicola and C. bolitinos to oral infection with eight serotypes of epizootic haemorrhagic disease virus
Authors: Paweska J. T. , Venter G. J. , Hamblin C.
Published: June 2005

Abstract.  The mechanisms involved in introduction, maintenance and perpetuation of epizootic haemorrhagic disease virus (EHDV) in South Africa are not fully understood. This paper reports on the susceptibility of South African livestock associated Culicoides (Diptera: Ceratopogonidae) species to oral infection with eight EHDV serotypes. Virus was recovered from eight of 17 field‐collected Culicoides species 10 days after oral feeding on blood/virus mixtures. Six EHDV serotypes were recovered from C. (Avaritia) imicola Kieffer, and seven serotypes were recovered from C. (A.) bolitinos Meiswinkel. Virus recovery rates in C. imicola ranged from 0.4% for EHDV 2 to 14.4% for EHDV 7, and in C. bolitinos from 0.6% for EHDV 6 to 12.3% for EHDV 2. There was a significant difference in virus recovery rates between serotypes in both species. Other Culicoides species that yielded EHDV after 10 days extrinsic incubation included C. (Meijerehelea) leucostictus Kieffer, C. (Culicoides) magnus Colaço, C. (Beltranmyia) nivosus de Meillon, C. (A.) gulbenkiani Caeiro, C. (Hoffmania) zuluensis de Meillon and C. onderstepoortensis Fiedler. Culicoides midges shown in this study to be susceptible to oral infection with EHDV are widely distributed in South Africa but differ considerably in their abundance, host preference and breeding sites.
Energy conservation: an alternative for investment in the oil sector for OPEC Member Countries
Authors: Zamani Mehrzad
Published: June 2005

Investment in the oil sector is the main policy of expanding net crude oil export capacity in OPEC Member Countries. The other alternative should be improving energy conservation policies. Since these countries benefit from cheap energy sources, it is reasonable to expect inefficient use of energy in their economies, resulting in relatively high energy intensity. This paper deals with the causality relationship between energy consumption and gross domestic product (GDP). First, stationary tests are run. Second, if there is a cointegrating relationship, an error correction model is applied; otherwise a standard Granger causality test is conducted. It was discovered that for all OPEC Member Countries we cannot statistically accept causality running from energy to GDP. Therefore, not only are proper conservation policies not a threat to economic growth, they also lead to an expansion of oil export capacity.
History of standard scoring, notation, and summation of neuromuscular signs. A current survey and recommendation
Authors: Dyck Peter J. , Boes Christopher J. , Mulder Donald , Millikan Clark , Windebank Anthony J. , Dyck P. James B. , Espinosa Raul
Published: June 2005

Abstract  In this article, we trace the history of scoring, notation, and summation of the neuromuscular signs of muscle weakness and decrease of tendon reflexes and sensation. We recommend a standard system to promote consistency in the effort introduced by Mitchell and Lewis to ‘represent systems and force by their signs.’ The scoring of neuromuscular signs began with Mitchell and Lewis in the 19th century who used pluses, minuses, and N (for normal) to express the activity of muscle stretch reflexes. Henry Plummer introduced an ordinal scoring approach for muscle weakness, reflex decrease and increase, and sensation loss. In 1919, he and Walter Sheldon and Henry Woltman introduced standard pre‐printed examination forms with written instructions for notation and scoring. Robert Lovett, a Boston orthopedist, scored weak muscles of poliomyelitis patients from 2 (mild weakness) to 6 (paralyzed), 1 being normal. Lovett's approach was used, after reversing the order of the grades and decreasing each grade by 1, by a Committee of the Medical Research Council for evaluating return of muscle weakness after nerve injury. Despite dissimilarity to existing reflex and sensation scores and uneven width of grades, this approach was widely adopted for use in neurologic practice. We introduced the Neuropathy Impairment Score using a combination of the Mitchell, Plummer, and Lovett approaches, summing all individual scores of a standard set of neuromuscular examinations. In a non‐representative survey of 19 neuromuscular physicians from different countries, we find that there is a considerable variability in the approaches used for grading. Assuming that scoring is useful, we herein suggest (a) impairments should be scored separately from hyperfunction and (b) for the scoring of impairments (muscle weakness, reflex decrease, and sensation loss), the same ordinal scoring approach should be used with 0 as normal and 1, 2, … representing increasing impairment based on the judgment of percentage abnormality with corrections made for age, sex, physical fitness, and physical characteristics.
Antibodies to peripheral nerve myelin proteins in chronic inflammatory demyelinating polyradiculoneuropathy
Authors: Allen David , Giannopoulos Konstantinos , Gray Ian , Gregson Norman , Makowska Anna , Pritchard Jane , Hughes Richard A. C.
Published: June 2005

Abstract  Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an acquired disorder of the peripheral nervous system with a probable auto‐immune pathogenesis. The nature of the responsible autoantigens is unclear in most patients. We used the Western immunoblot technique to seek antibodies to peripheral nerve protein antigens. Sera from eight of 32 (25%) CIDP patients, 12 of 37 (32%) Guillain‐Barré syndrome (GBS) patients, zero of 30 (0%) chronic idiopathic axonal polyneuropathy patients and two of 39 (5%) healthy control subjects contained anti‐peripheral nerve protein antibodies. The frequency of such antibodies was significantly greater in both CIDP (p = 0.04) and GBS (p = 0.003) patients than in normal control subjects. For CIDP patients, there were non‐significant trends for antibodies to be more common in females and in those who responded to treatment with either intravenous immunoglobulin or plasma exchange. The commonest antibodies were directed against a band at 28 kDa, resembling that labelled by a monoclonal antibody against myelin protein zero (P0). Six CIDP and seven GBS patients' sera reacted with this band. These results support the view that antibodies to myelin proteins, and especially P0, are present in the serum of some patients with CIDP and GBS.
Functional health status of patients with chronic inflammatory neuropathies
Authors: Erdmann Peter G. , Van Meeteren Nico L. U. , Kalmijn Sandra , Wokke John H. J. , Helders Paul J. M. , Van Den Berg Leonard H.
Published: June 2005

Abstract The functioning of 12 patients with chronic inflammatory demyelinating polyneuropathy (CIDP) and 18 patients with multifocal motor neuropathy (MMN) was evaluated to obtain health profiles and appropriate clinimetric instruments. Assessment was made in a cross‐sectional study by means of a performance‐based body function test (hand‐held dynamometry), two performance‐based activity tests (10‐m walk test and Berg balance scale), a self‐reported activity test (Canadian Occupational Performance Measure), and a self‐reported functioning test (sickness impact profile 68). In both patient groups, CIDP and MMN, specific health profiles were manifest. A clear relationship between body function, activities, and functioning was not found. Therefore, to assess a patient with inflammatory neuropathy, it is recommended to assess body function as well as activities and functioning and to select appropriate clinimetric instruments specific for each type of neuropathy.
Development and biological analysis of peritoneal metastasis mouse models for human scirrhous stomach cancer
Authors: Yanagihara Kazuyoshi , Takigahira Misato , Tanaka Hiromi , Komatsu Teruo , Fukumoto Hisao , Koizumi Fumiaki , Nishio Kazuto , Ochiya Takahiro , Ino Yoshinori , Hirohashi Setsuo
Published: June 2005

The number of published studies on peritoneal dissemination of scirrhous gastric carcinoma is very small as a result of the unavailability of highly reproducible animal models. Orthotopic implantation of HSC‐44PE and HSC‐58 (scirrhous gastric carcinoma‐derived cell lines) cells into nude mice led to dissemination of the tumor cells to the greater omentum, mesenterium, peritoneum and so on, and caused ascites in a small number of animals. Cycles of isolation of the ascitic tumor cells and orthotopic inoculation of these cells were repeated in turn to animals. This was to isolate highly metastatic cell lines with a strong capability of inducing the formation of ascites (44As3 from HSC‐44PE; 58As1 and 58As9 from HSC‐58). All three cell lines induced tumor formation at the site of orthotopic injection, and caused fatal cancerous peritonitis and bloody ascites in 90–100% of the animals approximately 3–5 weeks after the inoculation. When the parent cells were implanted, the animals became moribund in approximately 12–18 weeks, however, none of the animals developed ascites. Complementary DNA microarray and immunohistochemical analyses revealed differences in the expression levels of genes coding for the matrix proteinase, cell adhesion, motility, angiogenesis and proliferation between the highly metastatic‐ and parent‐cell lines. The usefulness of this model for the evaluation of drugs was assessed by analyzing the stability of the metastatic potential of the cells and the reproducibility. Animals intravenously treated with CPT‐11 and GEM showed suppressed tumor growth and significantly prolonged survival. The metastatic cell lines and the in vivo model established in the present study are expected to serve as a model of cancerous peritonitis developing from primary lesions, and as a useful means of clarifying the pathophysiology of peritoneal dissemination of scirrhous gastric carcinoma and the development of drugs for its treatment. (Cancer Sci 2005; 96: 323–332)
Inhibition of protein synthesis and activation of stress‐activated protein kinases by onnamide A and theopederin B, antitumor marine natural products
Authors: Lee Kun‐Hyung , Nishimura Shinichi , Matsunaga Shigeki , Fusetani Nobuhiro , Horinouchi Sueharu , Yoshida Minoru
Published: June 2005

During the course of screening for the agents that activate transforming growth factor‐β (TGF‐β) signaling cascade, onnamide A and theopederin B, heterocyclic compounds related to mycalamides from a marine sponge, were found to induce plasminogen activator inhibitor‐1 (PAI‐1) promoter‐derived gene expression in Mv1Lu cells. The maximum induction of the PAI‐1 promoter by onnamide A and theopederin B was observed at the concentrations of 50 nM and 2 nM, respectively. These compounds strongly inhibited protein synthesis at the 50% inhibitory concentrations of 30 nM for onnamide A and 1.9 nM for theopederin B, and induced activation of p38 mitogen‐activated protein kinase and c‐Jun NH2‐terminal protein kinase (JNK). Anisomycin, a well‐known inducer of ribotoxic stress that inhibits protein synthesis and activates both p38 kinase and JNK, also activated PAI‐1 gene expression. Furthermore, PAI‐1 expression by onnamide A, theopederin B, and anisomycin was inhibited by SB202190 and SP600125, specific inhibitors of stress‐activated protein kinases. Onnamide A and theopederin B were cytotoxic to a variety of cell lines and strongly induced apoptosis in HeLa cells within 24 h, which was accompanied by the sustained activation of p38 kinase and JNK. These results suggest that onnamide A and theopedirin B trigger a ribotoxic stress‐like response, thereby inducing p38 kinase and JNK activation, the kinase‐dependent PAI‐1 gene expression, and apoptosis. (Cancer Sci 2005; 96: 357–364)
T cell-mediated immune responses in human newborns: ready to learn?
Authors: Marchant A , Goldman M
Published: June 2005

Infections with intracellular pathogens are often more severe or more prolonged in young infants suggesting that T cell-mediated immune responses are different in early life. Whereas neonatal immune responses have been quite extensively studied in murine models, studies of T cell-mediated immunity in human newborns and infants are scarce. Qualitative and quantitative differences when compared with adult immune responses have been observed but on the other hand mature responses to certain vaccines and infectious pathogens were demonstrated during the postnatal period and even during foetal life. Herein, we review the evidence suggesting that under appropriate conditions of stimulation, protective T cell-mediated immune responses could be induced by vaccines in early life.
AcellularBordetella pertussisvaccine enhances mucosal interleukin-10 production, induces apoptosis of activated Th1 cells and attenuates colitis in Gαi2-deficient mice
Authors: Öhman L , Willén R , Hultgren O H , Hultgren Hörnquist E
Published: June 2005

Mice deficient for the inhibitory G protein subunit α2 (Gαi2–/–) spontaneously develop a progressive inflammatory bowel disease resembling ulcerative colitis, and have a T helper 1 (Th1)-dominated immune response prior to onset of colitis, which is further augmented after the onset of disease. The present study was performed to investigate whether the Gαi2–/– mice were able to down-regulate the Th1-dominated inflammatory mucosal immune response and/or induce an anti-inflammatory Th2/T regulatory response and thereby diminish the severity of colitis following treatment with acellular Bordetella pertussis vaccine. The acellular vaccine against B. pertussis, the causative agent of whooping cough, has been demonstrated to induce a Th2-mediated response in both man and mice. We therefore treated Gαi2–/– mice intraperitoneally with a three-component acellular B. pertussis vaccine. The treated Gαi2–/– mice showed significantly increased interleukin (IL)-10 production in intestinal tissue, associated with significantly reduced colitis and decreased mortality, compared to untreated Gαi2–/– mice. The attenuation of colitis in Gαi2–/– mice was due, at least partly, to the B. pertussis surface antigen filamentous haemagglutinin (FHA), which almost completely inhibited proliferation of CD4+ T cells and stimulated apoptosis of activated CD4+ T helper 1 cells. In conclusion, the three-component acellular B. pertussis vaccine containing filamentous haemagglutinin increases the production of IL-10 in the intestinal mucosa, induces apoptosis of activated Th1 cells and attenuates colitis in Gαi2–/– mice.
Cross-reaction between antibodies to the major epitope of Ro60 kD autoantigen and a homologous peptide of Coxsackie virus 2B protein
Authors: Stathopoulou E A , Routsias J G , Stea E A , Moutsopoulos H M , Tzioufas A G
Published: June 2005

Coxsackie virus RNA has recently been detected in biopsy specimens of minor salivary glands from patients with primary Sjögren's Syndrome (pSS). A peptide derived from Coxsackie virus 2B protein (pepCoxs) presents 87% sequence homology with the 222–229 region of the major linear B-cell epitope of  Ro60 kD  autoantigen  (pep216–232).  Synthetic  peptides  corresponding to pep216–232: 216KALSVETEKLLKYLEAV232 and pepCoxs: 31MVTSTITEKL LKNLVKI47, were prepared. Sera from 42 patients with pSS and 43 patients with systemic lupus erythematosus (SLE) as well as sera from 27 healthy individuals (normal controls) and sera from 30 patients with rheumatoid arthritis (disease controls) were tested against the two homologous peptides. Twenty-five percent of SLE sera and 33·3% of pSS sera reacted against pep216–232, whereas 28% of SLE sera and 37% of pSS sera recognized the pepCoxs. The sera reacting with pep216–232 were apparently the same as those reacting with pepCoxs. Normal sera and disease control sera presented only a limited reactivity against both peptides (ranging from 3·7% to 10%). Both peptides reacted more prominently with anti-Ro/La (+) sera from pSS patients. Thus, pep216–232 was recognized by 17% of the anti-Ro (+) sera and by 42% of the anti-Ro/La (+) sera, whereas pepCoxs was recognized by 28·5% and 38% of the a-Ro(+) and a-Ro/La(+) sera, respectively. Purified anti-pep216–232 antibodies readily reacted with both peptides while inhibition experiments revealed the specificity of this reaction. These results suggest a possible cross-reaction between antibodies to the major linear B-cell epitope of Ro60 kD autoantigen and the homologous pepCoxs in pSS patients. This cross-reaction might potentially play a role in autoantibody formation and the perpetuation of the autoimmune response against Ro/SSA and La/SSB.
Analysis of circulating γδ+ T cells in children affected by IgE-associated and non-IgE-associated allergic atopic eczema/dermatitis syndrome
Authors: Cairo C , Arabito E , Landi F , Casati A , Brunetti E , Mancino G , Galli E
Published: June 2005

Recent studies have suggested that not only αβ+ T cells, but also the less common γδ+ T cells may play a role as effectors and immunoregolatory cells in the development and perpetuation of allergic inflammation. The objective of this study was to focus on the role of γδ+ T cells in atopic dermatitis (AD), a chronic relapsing inflammatory disease of the skin, often associated with allergic bronchial asthma. The present study employed flow cytometric analysis to compare numbers and phenotypic characteristics of γδ+ T cells in the peripheral blood of children with atopic dermatitis and age-matched healthy controls. The percentage of circulating Vγ 9Vδ2+ T lymphocytes was significantly increased in AD patients with respect to the age-matched controls, with a positive correlation with clinical score severity. The prevalent phenotype in both AD patients and controls was CD45RO+, with no differences observed in the percentage of Vδ2+ CD45RO+ between these groups. Conversely, memory CD45RO+ CD62L+ Vδ2+ lymphocytes were significantly lower in AD patients. Furthermore, naive circulating Vδ2+ T lymphocytes were significantly lower in AD children than in aged-matched controls. No correlation was observed between circulating Vγ 9Vδ2+ expansion and IgE serum levels. It was concluded that an association exists between the levels of circulating γδ+ T lymphocytes and atopic dermatitis, with a positive correlation with clinical score but no link with IgE serum levels. The pathophysiological role of γδ T lymphocytes in atopic dermatitis awaits further investigation.
TCR Vα14+ natural killer T cells function as effector T cells in mice with collagen-induced arthritis
Authors: Ohnishi Y , Tsutsumi A , Goto D , Itoh S , Matsumoto I , Taniguchi M , Sumida T
Published: June 2005

Natural killer (NK) T cells are a unique, recently identified cell population and are suggested to act as regulatory cells in autoimmune disorders. In the present study, designed to investigate the role of NKT cells in arthritis development, we attempted to induce arthritis by immunization of type II collagen (CIA) in Jα281 knock out (NKT-KO) and CD1d knock out (CD1d-KO) mice, which are depleted of NKT cells. From the results, the incidence of arthritis (40%) and the arthritis score (1·5 ± 2·2 and 2·0 ± 2·7) were reduced in NKT-KO and CD1d-KO mice compared to those in respective wild type mice (90%, 5·4 ± 3·2 and 2·0 ± 2·7, P < 0·01). Anti-CII antibody levels in the sera of NKT-KO and CD1d-KO mice were significantly decreased compared to the controls (OD values; 0·32 ± 0·16 and 0·29 ± 0·06 versus 0·58 ± 0·08 and 0·38 ± 0·08, P < 0·01). These results suggest that NKT cells play a role as effector T cells in CIA. Although the cell proliferative response and cytokine production in NKT-KO mice after the primary immunization were comparable to those in wild type mice, the ratios of both activated T or B cells were lower in NKT-KO mice than wild type mice after secondary immunization (T cells: 9·9 ± 1·8%versus 16·0 ± 3·4%, P < 0·01, B cells: 4·1 ± 0·5%versus 5·1 ± 0·7%, P < 0·05), suggesting that inv-NKT cells contribute to the pathogenicity in the development phase of arthritis. In addition, IL-4 and IL-1β mRNA expression levels in the spleen during the arthritis development phase were lower in NKT-KO mice, while the IFN-γ mRNA expression level was temporarily higher. These results suggest that inv-NKT cells influence cytokine production in arthritis development. In conclusion, inv-NKT cells may promote the generation of arthritis, especially during the development rather than the initiation phase.
Increased monocyte transcription of the proteinase 3 gene in small vessel vasculitis
Authors: Ohlsson S , Hellmark T , Pieters K , Sturfelt G , Wieslander J , Segelmark M
Published: June 2005

Proteinase 3 (PR3) is a pleiotropic and destructive serine protease and it is also a major target for autoantibodies in systemic small vessel vasculitis. We have shown recently that patients in stable remission have increased circulating levels of PR3, independent of autoantibody titre, inflammation, neutrophil degranulation and renal function. Here we explore the possibility of increased PR3 gene transcription. RNA was purified from peripheral blood monocytes from vasculitis patients and controls. Specific mRNA was measured by TaqMan real-time polymerase chain reaction (PCR). The monocyte-like cell lines THP-1 and U937 and human peripheral blod monocytes from healthy controls were stimulated with cytokines and lipopolysaccharide (LPS) for different time periods. PR3 protein was measured in plasma with enzyme-linked immunosorbent assay (ELISA). The median result for PR3 mRNA was 9.6 (1.8–680) for 22 patients, compared to 1 (0.1–2.8) for the 15 healthy controls. Elastase expression was also significantly increased, whereas myeloperoxidase and interleukin-8 were not. Stimulation of monocytes with tumour necrosis factor (TNF)-α, interferon (IFN)-γ or LPS did not result in any increase of PR3 or elastase transcription, whereas interleukin (IL)-8 transcription was increased 10-fold. Circulating monocytes from patients with systemic vasculitis display increased PR3 gene transcription compared to healthy controls and patients with sytemic lupus erythematosus (SLE). This may be important for the development of vasculitis. Our results do not favour a role for cytokines, antineutrophil cytoplasmic antibodies (ANCA) or immunosuppressive medication in the upregulation of PR3 transcription in vasculitis.
Interleukin-4 induced down-regulation of skin homing receptor expression by human viral-specific CD8+ T cells may contribute to atopic risk of cutaneous infection
Authors: Seneviratne S L , Jones L , Bailey A S , Samuel R V , Black A P , Ogg G S
Published: June 2005

Factors controlling the expression of cutaneous lymphocyte-associated antigen (CLA) by T cells are poorly understood, but data from murine and human CD4+ T cell systems have suggested that cytokines play an important role. However, there are no data examining the influence of cytokines on the expression of CLA by human antigen-specific CD8+ T cells. Peripheral blood mononuclear cells (PBMC) were isolated from 10 HLA-A*0201-positive healthy individuals. Using HLA-peptide tetrameric complexes refolded with immunodominant peptides from Epstein–Barr virus (EBV), cytomegalovirus (CMV) and influenza A virus, we investigated the temporal associations of CLA expression by viral-specific CD8+ T cells following stimulation with antigen. Ex vivo influenza matrix-specific CD8+ T cells expressed significantly (P < 0·05) greater levels of CLA than EBV BMLF1 and CMV pp65-specific CD8+ T cells (mean 9·7% influenza matrix versus 1·4% BMLF1 versus 1·1% pp65) and these differences were sustained on culture. However, regardless of viral specificity, interleukin (IL)-12 and IL-4 induced significant (P < 0·05) dose-dependent up-regulation and down-regulation of CLA expression, respectively, with IL-4 showing a dominant negative effect. In many cases, IL-4 resulted in complete abrogation of detectable CLA expression by the viral-specific CD8+ T cells. Overall these data demonstrate that CLA expression by human viral-specific CD8+ T cells is highly dynamic and that IL-4 causes significant down-regulation. Disorders associated with a type 2 cytokine shift may reduce the efficiency of skin homing by viral-specific CD8+ T cells. Furthermore, the ability to modify the local and systemic microenvironment may offer novel therapeutic strategies that influence tissue-specific T cell homing.