bluuebunny/crossref_metadata_2025 huggingface dataset:
H5N1 influenza virus evolution: a comparison of different epidemics in birds and humans (1997–2004)
Authors: Campitelli Laura , Ciccozzi Massimo , Salemi Marco , Taglia Fabiana , Boros Stefano , Donatelli Isabella , Rezza Giovanni
Published: March 2006

The selection pressure acting along the entire genome sequence of H5N1 avian influenza viruses isolated from several bird species and humans infected in the 1997 and 2004 outbreaks, and on the HA1 genes from H5N1 viruses isolated during the entire study period, in eastern Asia was evaluated. According to maximum-likelihood analysis, viral genes appeared to be, in both epidemics, under strong purifying selection, with only the PB2, HA and NS1 genes under positive selection. Specific codons under positive selection were detected by using codon-based substitution models. Positive-selection analysis performed on single-codon sites might be helpful in clarifying the driving force of avian and human influenza virus evolution and in selecting specific targets for vaccines and antiviral drugs.
Deletions in the hypervariable domain of the nsP3 gene attenuate Semliki Forest virus virulence
Authors: Galbraith Sareen E. , Sheahan Brian J. , Atkins Gregory J.
Published: March 2006

Major virulence determinants ofSemliki Forest virus(SFV) lie within the non-structural genes that form the replicase complex proteins. Gene exchange between virulent and avirulent viruses has shown that the nsP3 gene, which has essential 5′ conserved domains and a non-essential hypervariable 3′ domain, is one of the virulence determinants. This protein plays a role in subgenomic 26S and negative-strand RNA synthesis and is thought to function with nsP1 to anchor replication complexes to cell membrane structures. Studies to date have focused on analysing the effect of mutational changes spread over the whole gene on virulence of the virus. The virulent SFV4 virus, derived from an infectious clone, was utilized to analyse the effect on virulence of large deletions in the hypervariable domain of nsP3. Two viruses with different in-frame deletions that spanned this domain showed reduced rates of RNA synthesis and multiplication in cell culture. In adult BALB/c mice, these viruses were avirulent after intramuscular and intraperitoneal inoculation, and brains sampled from infected mice showed minimal or no evidence of pathology. These deleted viruses had greatly reduced virulence when administered by the intranasal route and brains from infected mice showed lesions that were much less severe than those seen in SFV4 infection. Mice surviving infection with the deleted viruses resisted challenge with the virulent L10 strain, indicating induction of protective immunity. This work establishes that deletions in the nsP3 hypervariable domain attenuate virulence after peripheral inoculation and also reduce virulence after intranasal inoculation.
Yaba-like disease virus chemokine receptor 7L, a CCR8 orthologue
Authors: Najarro Pilar , Gubser Caroline , Hollinshead Michael , Fox James , Pease James , Smith Geoffrey L.
Published: March 2006

Yaba-like disease virus (YLDV) gene7Lencodes a seven-transmembrane G protein-coupled receptor with 53 % amino acid identity to human CC chemokine receptor 8 (CCR8). Initial characterization of 7L showed that this 56 kDa cell-surface glycoprotein binds human CCL1 with high affinity (Kd=0·6 nM) and induces signal transduction by activation of heterotrimeric G proteins and downstream protein kinases. Further characterization of YLDV 7L is presented here and shows that murine CC chemokines can induce G-protein activation via the 7L receptor, despite having a low binding affinity for this receptor. In addition, when expressed by recombinant vaccinia virus (VACV), YLDV 7L was found on the outer envelope of VACV extracellular enveloped virus. The contribution of 7L to poxvirus pathogenesis was investigated by infection of mice with a recombinant VACV expressing 7L (vΔB8R-7L) and was compared with the outcome of infection by parental and revertant control viruses. In both intranasal and intradermal models, expression of 7L caused attenuation of VACV. The role of this protein in viral virulence is discussed.
Identification of prion protein gene polymorphisms in goats from Italian scrapie outbreaks
Authors: Acutis P. L. , Bossers A. , Priem J. , Riina M. V. , Peletto S. , Mazza M. , Casalone C. , Forloni G. , Ru G. , Caramelli M.
Published: March 2006

Susceptibility to scrapie in sheep is influenced by polymorphisms of the prion protein (PrP) gene, whereas no strong association between genetics and scrapie has yet been determined in goats due to the limited number of studies on these animals. In this case–control study on 177 goats from six Italian scrapie outbreaks, the association between PrP alleles and the occurrence of scrapie was studied. Three silent mutations and 11 PrP polymorphisms were identified, of which two polymorphisms (L133Q and M137I) and one silent mutation (T202T) have not been reported previously. Twelve alleles were determined by cloning. Statistical analysis suggested a possible protective role against scrapie for the glutamine to lysine mutation at codon 222.
Quantification of Feline immunodeficiency virus (FIVpco) in peripheral blood mononuclear cells, lymph nodes and plasma of naturally infected cougars
Authors: Blake David J. , Graham Jon , Poss Mary
Published: March 2006

Infection of domestic cats with Feline immunodeficiency virus (FIV) results in a fatal immunodeficiency disease, similar to Human immunodeficiency virus 1 (HIV-1) in humans. Elevated plasma viral loads in domestic cats are correlated to decreased survival time and disease progression. However, FIV is also maintained as an apathogenic infection in other members of the family Felidae including cougars, Puma concolor (FIVpco). It is not known whether the lack of disease in cougars is a result of diminished virus replication. A real-time PCR assay was developed to quantify both FIVpco proviral and plasma viral loads in naturally infected cougars. Proviral loads quantified from peripheral blood mononuclear cells (PBMC) ranged from 2·90×101 to 6·72×104 copies per 106 cells. Plasma viral loads ranged from 2·30×103 to 2·81×106 RNA copies ml−1. These data indicate that FIVpco viral loads are comparable to viral loads observed in endemic and epidemic lentivirus infections. Thus, the lack of disease in cougars is not due to low levels of virus replication. Moreover, significant differences observed among cougar PBMC proviral loads correlated to viral lineage and cougar age (P=0·014), which suggests that separate life strategies exist within FIVpco lineages. This is the first study to demonstrate that an interaction of lentivirus lineage and host age significantly effect proviral loads.
Replication-dependent fitness recovery of Human immunodeficiency virus 1 harbouring mutations of Asn17 of the nucleocapsid protein
Authors: Tözsér József , Shulenin Sergey , Young Matthew R. , Briggs Carlton J. , Oroszlan Stephen
Published: March 2006

The genetic stability of attenuated Human immunodeficiency virus 1 (HIV-1) variants harbouring mutations (Gly or Lys) of Asn17, the protease-cleavage site of the proximal zinc finger of the nucleocapsid protein, was studied. All possible codons for the Gly mutants were tested as starting sequences. Long-term replication assays revealed that the mutants were unstable; mutations of Gly17 to Arg, Ala, Ser and Cys, as well as a Lys17Asn reversion, were observed. Replication kinetic assays in H9 cells revealed that the replication of Ala, Ser and Arg mutants was improved substantially compared with the Gly variant; the infectivity of Ala17 and Ser17 viruses was equal to, and that of Arg17 was almost equal to, the infectivity of the wild-type virus. Kinetic analysis of the cleavage of oligopeptides representing the corresponding nucleocapsid-cleavage sites revealed that all mutations improved cleavability, in good agreement with the previously proposed role of nucleocapsid cleavage in HIV-1 replication.
Isolation of a new strain of the flavivirus cell fusing agent virus in a natural mosquito population from Puerto Rico
Authors: Cook Shelley , Bennett Shannon N. , Holmes Edward C. , De Chesse Reine , Moureau Gregory , de Lamballerie Xavier
Published: March 2006

The genus Flavivirus contains approximately 70 single-stranded, positive-sense RNA viruses that are mosquito-borne, tick-borne or have no known vector. Two discoveries support previous suggestions of the existence of a large number of unsampled flaviviruses: (i) a new flavivirus, Kamiti River virus, was recently isolated from Kenyan mosquitoes, and (ii) sequences with high similarity to those of flaviviruses have been found integrated into the genome of Aedes mosquitoes, suggesting a past infection with a virus (or viruses) that has yet to be discovered. These sequences were related most closely to a flavivirus that infects insects alone, cell fusing agent virus (CFAV). CFAV was originally isolated in the laboratory from an Aedes aegypti cell line. To date, this virus had not been found in the wild. In the present study, over 40 isolates of a novel strain of CFAV were discovered from mature mosquitoes sampled from the wild in Puerto Rico. The viral strain was present in a range of mosquito species, including Aedes aegypti, Aedes albopictus and Culex sp., from numerous locations across the island and, importantly, in mosquitoes of both sexes, suggesting vertical transmission. Here, results from viral screening, and cell culture and molecular identification of the infected mosquitoes are presented. Experimental-infection tests were also conducted by using the original CFAV strain and a highly efficient reverse-transcription mechanism has been documented, in which initiation of copying occurs at the 3′ terminus of either the genomic RNA or the intermediate of replication, potentially elucidating the mechanism by which flaviviral sequences may have integrated into mosquito genomes.
Antiviral effect of α-glucosidase inhibitors on viral morphogenesis and binding properties of hepatitis C virus-like particles
Authors: Chapel Cynthia , Garcia Céline , Roingeard Philippe , Zitzmann Nicole , Dubuisson Jean , Dwek Raymond A. , Trépo Christian , Zoulim Fabien , Durantel David
Published: March 2006

Hepatitis C virus (HCV) infections are a major public-health concern. New antiviral drugs are needed urgently to complement and improve the efficacy of current chemotherapies. The morphogenesis of HCV represents an interesting, and still unexploited, novel molecular target.α-Glucosidase inhibitors derived from the glucose analogue deoxynojirimycin (DNJ) inhibit viral morphogenesisin cellulovia perturbation of theN-glycosylation pathway and hence the misfolding of viral glycoproteins that depend on certainN-glycans for correct folding. Due to the heavyN-glycosylation of HCV glycoproteins, it was hypothesized that such inhibitors would also affect HCV morphogenesis. To study the effect ofα-glucosidase inhibitors on viral morphogenesis and binding properties, HCV virus-like particles (VLPs) were produced by using baculovirus loaded with HCV structural-protein genes. Here, it is demonstrated that, in the presence of theseα-glucosidase inhibitors, viral glycoproteins synthesized and retained in the endoplasmic reticulum (i) contain unprocessed, triglucosylatedN-glycans, (ii) are impaired in their interaction with calnexin and (iii) are at least partially misfolded. Moreover, it is shown that, although the production of VLPs is not affected byα-glucosidase inhibitors, these VLPs contain unprocessed, triglucosylatedN-glycans and potentially misfolded glycoproteins. Finally, it is demonstrated that VLPs produced in the presence ofα-glucosidase inhibitors have impaired binding properties to hepatoma cells. The inhibitors of morphogenesis studied here target steps of the HCV viral cycle that may prevent or delay viral resistance. Theseα-glucosidase inhibitors may prove to be useful molecules to fight HCV infection in combination protocols.
Characterization and immunological analysis of the rhesus cytomegalovirus homologue (Rh112) of the human cytomegalovirus UL83 lower matrix phosphoprotein (pp65)
Authors: Yue Yujuan , Kaur Amitinder , Zhou Shan Shan , Barry Peter A.
Published: March 2006

Rhesus cytomegalovirus (RhCMV) contains two open reading frames (Rh111 and Rh112) that encode proteins homologous to the phosphoprotein 65 (pp65) of the human cytomegalovirus (HCMV) UL83 gene. As HCMV pp65 elicits protective immune responses in infected humans and represents an important vaccination target, one RhCMV homologue of HCMV pp65, pp65-2 (Rh112), was characterized and analysed for its ability to induce host immune responses. Similar to its HCMV counterpart, RhCMV pp65-2 was expressed as a late gene, localized to the nucleus within pp65-2-expressing cells and was present within infectious virions. Longitudinal and cross-sectional studies of pp65-2 immunity in naturally infected rhesus macaques showed that humoral responses to pp65-2 were elicited early during infection, but were not always sustained over time. In contrast, pp65-2-specific T-cell responses, examined by gamma interferon ELISPOT, were broadly detectable in all of the animals studied during primary infection and persisted in the vast majority of RhCMV-seropositive monkeys. Moreover, there was considerable inter-animal variability in the pattern of the immune responses to pp65-2. Together, these results demonstrated that RhCMV pp65-2 exhibited biological and immunological homology to HCMV pp65. Thus, the rhesus macaque model of HCMV persistence and pathogenesis should be relevant for addressing pp65-based vaccine modalities.
Genetic and antigenic diversity among noroviruses
Authors: Hansman Grant S. , Natori Katsuro , Shirato-Horikoshi Haruko , Ogawa Satoko , Oka Tomoichiro , Katayama Kazuhiko , Tanaka Tomoyuki , Miyoshi Tatsuya , Sakae Kenji , Kobayashi Shinichi , Shinohara Michiyo , Uchida Kazue , Sakurai Nakao , Shinozaki Kuniko , Okada Mineyuki , Seto Yoshiyuki , Kamata Kunio , Nagata Noriyo , Tanaka Keiko , Miyamura Tatsuo , Takeda Naokazu
Published: March 2006

Human norovirus (NoV) strains cause a considerable number of outbreaks of gastroenteritis worldwide. Based on their capsid gene (VP1) sequence, human NoV strains can be grouped into two genogroups (GI and GII) and at least 14 GI and 17 GII genotypes (GI/1–14 and GII/1–17). Human NoV strains cannot be propagated in cell-culture systems, but expression of recombinant VP1 in insect cells results in the formation of virus-like particles (VLPs). In order to understand NoV antigenic relationships better, cross-reactivity among 26 different NoV VLPs was analysed. Phylogenetic analyses grouped these NoV strains into six GI and 12 GII genotypes. An antibody ELISA using polyclonal antisera raised against these VLPs was used to determine cross-reactivity. Antisera reacted strongly with homologous VLPs; however, a number of novel cross-reactivities among different genotypes was observed. For example, GI/11 antiserum showed a broad-range cross-reactivity, detecting two GI and 10 GII genotypes. Likewise, GII/1, GII/10 and GII/12 antisera showed a broad-range cross-reactivity, detecting several other distinct GII genotypes. Alignment of VP1 amino acid sequences suggested that these broad-range cross-reactivities were due to conserved amino acid residues located within the shell and/or P1-1 domains. However, unusual cross-reactivities among different GII/3 antisera were found, with the results indicating that both conserved amino acid residues and VP1 secondary structures influence antigenicity.
Hepatitis C virus-related internal ribosome entry sites are found in multiple genera of the family Picornaviridae
Authors: Chard Louisa S. , Bordeleau Marie-Eve , Pelletier Jerry , Tanaka Junichi , Belsham Graham J.
Published: March 2006

The internal ribosome entry site (IRES) elements from porcine enterovirus 8 and simian virus 2, two members of a proposed new genus within the familyPicornaviridae, were characterized. These IRES elements, in common with the porcine teschovirus 1 IRES, were found to be related functionally and structurally to the IRES element fromHepatitis C virus, a member of the familyFlaviviridae. Partial secondary structure predictions were derived and functional assays demonstrated that these IRES elements continued to be active when eIF4G was cleaved and when the activity of eIF4A was blocked.
Phylogeny, recombination and expression of porcine endogenous retrovirus γ2 nucleotide sequences
Authors: Klymiuk Nikolai , Müller Mathias , Brem Gottfried , Aigner Bernhard
Published: March 2006

Endogenous retroviral sequences in the pig genome represent a potential infectious risk in xenotransplantation. Porcine endogenous retrovirus (PERV) γ sequences described to date have been classified into several families. The known infectious, human-tropic PERVs have been assigned to the PERV γ1 subfamilies A, B and C. High copy numbers and full-length clones have also been observed for an additional family, designated PERV γ2. The aim of this study was to examine the PERV γ2 family by analysis of retroviral pro/pol gene sequences. The proviral load was observed to be similar among various pig breeds. Although clones harbouring an open reading frame in the examined region were found, analysis of published large PERV γ2 clones revealed multiple deleterious mutations in each of the retroviral genes. Various recombination events between γ2 genomes were revealed. In contrast to PERV γ1, phylogenetic analyses did not distinguish defined subfamilies, but indicated the independent evolution of the proviruses after a single event of retroviral amplification. Expression analysis showed large PERV γ2 transcripts and variable transcription in several tissues. Analysis of the two published γ2 env gene sequences observed the partial lack of the receptor-binding domain. Overall, this study indicated the low infectious potential for PERV γ2.
The M4 gene of murine gammaherpesvirus 68 modulates latent infection
Authors: Geere Hannah M. , Ligertwood Yvonne , Templeton Kerra M. , Bennet Ian , Gangadharan Babunilayam , Rhind Susan M. , Nash Anthony A. , Dutia Bernadette M.
Published: March 2006

Murine gammaherpesvirus 68 (MHV-68) encodes a set of unique genes, M1, M2, M3 and M4, and eight non-translated tRNA-like molecules that are thought to be important in virus–host interactions and latent infection. The M4 gene is predicted to encode a novel secreted protein. To investigate the role of M4 in viral pathogenesis, a mutant MHV-68 that did not express M4 was constructed and its replication was characterized in vitro and in vivo. Virus replication was identical to the wild type in vitro and no difference could be detected in virus replication in the lung following intranasal infection. However, in the spleen, virus deficient in M4 expression was severely attenuated in the establishment of latency. These results indicate a critical role for M4 in MHV-68 pathogenesis.
Function, oligomerization and N-linked glycosylation of the Helicoverpa armigera single nucleopolyhedrovirus envelope fusion protein
Authors: Long Gang , Westenberg Marcel , Wang Hualin , Vlak Just M. , Hu Zhihong
Published: March 2006

In the familyBaculoviridae, two distinct envelope fusion proteins are identified in budded virions (BVs). GP64 is the major envelope fusion protein of group I nucleopolyhedrovirus (NPV) BVs. An unrelated type of envelope fusion protein, named F, is encoded by group II NPVs. The genome ofHelicoverpa armigera(Hear) NPV, a group II NPV of the single nucleocapsid or S type, also encodes an F-like protein: open reading frame 133 (Ha133). It was demonstrated by N-terminal sequencing of the major 59 kDa protein present in HearNPV BV that this protein is one of the two F subunits: F1(transmembrane subunit of 59 kDa) and F2(surface subunit of 20 kDa), both the result of cleavage by a proprotein convertase and disulfide-linked. The HearNPV F protein proved to be a functional analogue of GP64, as the infectivity of an AcMNPVgp64-deletion mutant was rescued by the introduction of the HearNPV F gene. It was also demonstrated by chemical cross-linking that HearNPV F is present in BVs as an oligomer whereby, unlike GP64, disulfide bonds are not involved. Deglycosylation assays indicated that both F1and F2possessN-linked glycans. However, when F was made in Hz2E5 cells, these glycans did not have anα-1-3 core fucose modification that usually occurs in insect cells. Asα-1-3 core fucose is a major inducer of an allergic response in humans, the present observation makes the HearNPV–Hz2E5 system an attractive alternative for the production of recombinant glycoproteins for therapeutic use in humans.
An essential role of ERK signalling in TPA-induced reactivation of Kaposi's sarcoma-associated herpesvirus
Authors: Cohen Adina , Brodie Chaya , Sarid Ronit
Published: March 2006

Kaposi's sarcoma-associated herpesvirus (KSHV) is implicated causally in the development of several human malignancies, including primary effusion lymphoma (PEL). PEL cells serve as tools for KSHV research, as most of them are latently infected and allow lytic virus replication in response to various stimuli. 12-O-Tetradecanoyl-phorbol-13-acetate (TPA) is the most potent inducer of lytic KSHV reactivation; nevertheless, the exact mechanism by which it induces reactivation remains unknown. It has previously been reported by our group that the protein kinase C (PKC)δisoform plays a crucial role in TPA-mediated KSHV reactivation. Here, the activation pathway was dissected and it was demonstrated that TPA induces KSHV reactivation via stimulation of the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway. Western blot analysis revealed a rapid phosphorylation of ERK1/2. Cells treated with MAPK/ERK inhibitors before TPA addition demonstrated repression of ERK1/2 phosphorylation, which was associated with a block of KSHV lytic-gene expression. This inhibition prevented c-Fos accumulation, yet increased c-Jun phosphorylation. Similar results were obtained in response to rottlerin, a selective PKCδinhibitor. Notably, the PKC inhibitor GF 109203X reduced ERK1/2 phosphorylation, c-Fos accumulation, c-Jun phosphorylation and KSHV reactivation. It is proposed that TPA induces KSHV reactivation through at least two arms. The first involves PKCδ, ERK phosphorylation and c-Fos accumulation, whilst the second requires another PKC isoform that induces the phosphorylation of c-Jun. c-Fos and c-Jun jointly form an active AP-1 complex, which functions to activate the lytic cascade of KSHV.
Corchorus yellow vein virus, a New World geminivirus from the Old World
Authors: Ha Cuong , Coombs Steven , Revill Peter , Harding Rob , Vu Man , Dale James
Published: March 2006

A bipartite begomovirus infecting Jute mallow (Corchorus capsularis, Tilliaceae) in Vietnam was identified using novel degenerate PCR primers. Analysis of this virus, which was named Corchorus yellow vein virus (CoYVV), showed that it was more similar to New World begomoviruses than to viruses from the Old World. This was based on the absence of an AV2 open reading frame, the presence of an N-terminal PWRLMAGT motif in the coat protein and phylogenetic analysis of the DNA A and DNA B nucleotide and deduced amino acid sequences. Evidence is provided that CoYVV is probably indigenous to the region and may be the remnant of a previous population of New World begomoviruses in the Old World.
Nuclear localization of the Epstein–Barr virus EBNA3B protein
Authors: Burgess Anita , Buck Marion , Krauer Kenia , Sculley Tom
Published: March 2006

The Epstein–Barr virus nuclear antigen (EBNA) 3B is a hydrophilic, proline-rich, charged protein that is thought to be involved in transcriptional regulation and is targeted exclusively to the cell nucleus, where it localizes to discrete subnuclear granules. Co-localization studies utilizing a fusion protein between enhanced green fluorescent protein (EGFP) and EBNA3B with FLAG-tagged EBNA3A and EBNA3C proteins demonstrated that EBNA3B co-localized with both EBNA3A and EBNA3C in the nuclei of cells when overexpressed. Computer analyses identified four potential nuclear-localization signals (NLSs) in the EBNA3B amino acid sequence. By utilizing fusion proteins with EGFP, deletion constructs of EBNA3B and site-directed mutagenesis, three of the four NLSs (aa 160–166, 430–434 and 867–873) were shown to be functional in truncated forms of EBNA3B, whilst an additional NLS (aa 243–246) was identified within the N-terminal region of EBNA3B. Only two of the NLSs were found to be functional in the context of the full-length EBNA3B protein.
Molecular tracing of Japan-indigenous hepatitis E viruses
Authors: Tanaka Yasuhito , Takahashi Kazuaki , Orito Etsuro , Karino Yoshiyasu , Kang Jong-Hon , Suzuki Kazuyuki , Matsui Atsushi , Hori Akiko , Matsuda Hiroyuki , Sakugawa Hiroshi , Asahina Yasuhiro , Kitamura Tsuneo , Mizokami Masashi , Mishiro Shunji
Published: March 2006

The ancestor(s) of apparently Japan-indigenous strains of Hepatitis E virus (HEV) was probably of foreign origin, but it remains unclear when and from where it made inroads. In this study, 24 genotype 3 and 24 genotype 4 HEV strains recovered in Japan each showed a significant cluster, clearly distinct from those of foreign strains, in the phylogenetic tree constructed from an 821 nt RNA polymerase gene fragment. The evolutionary rate, approximately 0·8×10−3 nucleotide substitutions per site per year, enabled tracing of the demographic history of HEV and suggested that the ancestors of Japan-indigenous HEV had made inroads around 1900, when several kinds of Yorkshire pig were imported from the UK to Japan. Interestingly, the evolutionary growth of genotype 3 in Japan has been slow since the 1920s, whereas genotype 4 has spread rapidly since the 1980s. In conclusion, these data suggest that the indigenization and spread of HEV in Japan were associated with the popularization of eating pork.
Core protein cleavage by signal peptide peptidase is required for hepatitis C virus-like particle assembly
Authors: Ait-Goughoulte Malika , Hourioux Christophe , Patient Romuald , Trassard Sylvie , Brand Denys , Roingeard Philippe
Published: March 2006

Hepatitis C virus (HCV) core protein, expressed with a Semliki Forest virus replicon, self-assembles into HCV-like particles (HCV-LP) at the endoplasmic reticulum (ER) membrane, providing an opportunity to study HCV assembly and morphogenesis by electron microscopy. This model was used to investigate whether the processing of the HCV core protein by the signal peptide peptidase (SPP) is required for the HCV-LP assembly. Several mutants were designed as there are conflicting reports concerning the cleavage of mutant proteins by SPP. Production of the only core mutant protein that escaped SPP processing led to the formation of multiple layers of electron-dense ER membrane, with no evidence of HCV-LP assembly. These data shed light on the HCV core residues involved in SPP cleavage and suggest that this cleavage is essential for HCV assembly.
Characterization of white spot syndrome virus replication in in vitro-cultured haematopoietic stem cells of freshwater crayfish, Pacifastacus leniusculus
Authors: Jiravanichpaisal Pikul , Söderhäll Kenneth , Söderhäll Irene
Published: March 2006

Replication of White spot syndrome virus (WSSV) was investigated in haematopoietic cells (hpt cells) derived from haematopoietic tissue (hpt) of freshwater crayfish, Pacifastacus leniusculus. Temperature and type of inoculum for virus replication were studied. The cell culture remained viable at a wide range of temperatures ranging from 4 to 25 °C. WSSV replicated in cells, as evidenced by in situ hybridization, RT-PCR and by the presence of virions visualized with an electron microscope. Moreover, the results showed that the infectivity of WSSV to hpt cells is dependent on temperature and a supplemented growth factor (cytokine) astakine. WSSV replicated more rapidly at higher temperatures than at lower temperatures. No virus replication was observed at 4 °C. Detectable WSSV-infected cells were present as early as 36 h post-inoculation, demonstrated by in situ hybridization or RT-PCR of VP28 expression at 25 °C. Hpt cells can survive a few weeks at 25 or 16 °C and longer than several months at 4 °C.
From ‘Vergangenheitsbewältigung’ to Generational Memory Contests in Günter Grass, Monika Maron and Uwe Timm
Authors: Fuchs Anne
Published: March 2006

Against the backdrop of the proliferation of trauma theory in public and academic debates, the paper argues that in spite of a recent trend to question the canonised discourse of contrition, German discussions about the legacy of the Nazi past are still largely dominated by the following two paradigms: a first psychoanalytic narrative tells a drama of repression, acting out and repetition compulsion, and a second narrative then restores the Enlightenment paradigm through its story of critical engagement with the past. In order to avoid the pitfall of an unhelpful demonisation of forgetting on the one hand, and the danger of a displaced sacralisation of the Holocaust on the other, the paper introduces the notion of memory contests into the debate. Provisionally defined as retrospective imaginings that simultaneously articulate, question and investigate the normative self‐image of previous generations, this notion accounts for the intergenerational dynamic of recent German identity debates. The dynamic of post‐‘Wende’ memory contests is explored with reference to recent works by Grass, Maron and Timm.
On the (In)Compatibility of Guilt and Suffering in German Memory1
Authors: Assmann Aleida
Published: March 2006

This article analyses the current shift in German memory concerning the issue of German suffering at the end of the Second World War. Contrary to widely held belief, these themes are not novel: German suffering was a topic of discourse immediately after the war in the private and political sphere. What is new in the current context, however, is the intensity of the unexpected return of these issues and their wide social resonance among different classes and generations. With this shift in focus, new memory contests arise. One paradigmatic case is the polarity created between a memory of German guilt and a memory of German suffering as represented by the two popular historians Hannes Heer and Jörg Friedrich; another concerns the (still ongoing) debate around a new centre for flight and expulsion. It is argued that the impasse of recent cultural memory debate typified by Heer and Friedrich can be surpassed by a more complex understanding of the structure of memory. According to this view, various levels of heterogeneous memory can exist side by side if they are contained within a normative frame of generally accepted validity.
Talkin’’Bout My Generation :Memories of 1968 in Recent German Novels
Authors: Shafi Monika
Published: March 2006

This article examines the depiction of 1968 in the novel Rot (2001) by Uwe Timm, in the narrative Mein Jahrhundert (1999) by Günter Grass, and in the autobiographical novel Die Brücke vom goldenen Horn (1998) by Emine Sevgi Özdamar, asking to what extent the concept of generation, understood sociologically and symbolically, is useful in analysing West Germany's 1968 generation and its legacy. The three authors display not only contrasting generational, literary and political profiles, they also entertain a different relationship to German mainstream culture. It becomes clear that Özdamar's novel unsettles precisely this dichotomy between the German mainstream and a multicultural niche‐discourse in its intense engagement with the 1968 movement in Germany and Europe. Her text therefore invites us to reconsider the value of the generational parameter in assessing the events of 1968.
Melancholy Competitions: W.G. Sebald Reads Günter Grass and Wolfgang Hildesheimer
Authors: Cosgrove Mary
Published: March 2006

This article examines how W.G. Sebald's melancholy understanding of history influences his reading of Günter Grass's Aus dem Tagebuch einer Schnecke und Wolfgang Hildesheimer's Tynset. Sebald pits Grass against Hildesheimer in a melancholy competition which tests the authenticity of 1960s discourse on ‘Trauer’. His criteria for the standard of authentic sadness are obscured, however, by his analysis of the writers’ use of symbols and references from Western writing on melancholy. Going beyond this surface of literary criticism, the article finds that Sebald's understanding and representation of history as melancholy is motivated by complexities of generational affiliation, ethnic identity and historical absenteeism. It is argued that Sebald's preference for what Dominick LaCapra terms the ‘transhistorical’ overlaps with aspects of American trauma theory, and that this tendency, alongside his critique of postwar Germany and his identification with the Jewish victims of history, accounts for his literary judgment in this instance. The article concludes that Sebald's strategy of argumentation favours an understanding of ‘Trauer’ as terminal melancholy, and further, that this register, as distinct from modes of mourning and ‘working‐through’, is the only acceptable discourse in which to commemorate Jewish victims of the Holocaust.
‘Memory Metonymies’: Music and Photography in Ingeborg Bachmann and Monika Maron
Authors: Ben‐Horin Michal
Published: March 2006

This article examines two novels that deal with the problem of past representation in German literature: Malina (1971) by Ingeborg Bachmann and Pawels Briefe (1999) by Monika Maron. These novels incorporate either musical or photographic images in a metonymic mode that forms an alternative poetics of memory. Metonymy is a figure of speech in which a part implicitly represents the whole, thus not constituting the ‘thing’ but rather hinting at it. In shaping acoustic as well as visual metonymic manifestations, both Bachmann and Maron create poetics that reveal the partial, fragmented nature of memory regarding the ‘catastrophe’ in Europe. However, the transformation from voice to picture and from anonymous subjects to private family narratives, which might be associated with ‘normalisation’ processes, also indicates a change in dealing with the past, as it is reflected in post‐unification literature.
History, Memory and Montage in Anne Duden's Das Judasschaf
Authors: Ludden Teresa
Published: March 2006

This article uses Theodor Adorno's ideas on the role of the artwork and Walter Benjamin's philosophy of history and his concept of ‘Eingedenken’ to examine the types of memory and the modes of remembering the Holocaust in Anne Duden's novel, Das Judasschaf (1985). It focuses on the complex re‐figuring of quotations, images, Renaissance paintings, and documentary materials in order to assess the politics of montage and the presenting of documents which interrupt the flow of the narrative. Analysis of the use of some contemporary historiographical concepts such as identification, empathy and ‘the gap’ feeds into the reading of Duden's text. The argument centres on non‐empathetic remembrance rather than identification with the victims as a more useful way of understanding the types of memory in the text.
The Ethics of Post‐Holocaust Art: Reflections on Redemption and Representation
Authors: Cooke Maeve
Published: March 2006

The article deals with ethical questions relating to artistic responses to catastrophe. Its starting point is James E. Young's thesis that post‐Holocaust art must renounce the traditional redemptory promises of art. Against this, drawing on the work of Theodor W. Adorno, it makes the case for art's redemptory dimension. Whereas Young and Adorno disagree with regard to the idea of redemption, they agree in their approval of a certain kind of negative aesthetics. Both writers advocate particular kinds of art as the ethically most appropriate response to catastrophe. The article raises questions concerning the ethical dimension of the kinds of art they advocate. In addition, it warns against hypostasising just one kind of art as the only ethically valid response to catastrophe. Finally, it argues that representational works of art are also important since they offer a particular ethical perspective that can stimulate reflection and debate.
Remembering the Berlin Wall: The Wall Memorial Ensemble Bernauer Strasse
Authors: Knischewski and Gerd , Spittler Ulla
Published: March 2006

Remembrance and commemoration of the National Socialist past have always been contentious issues within the political culture of West Germany. After unification this process of ‘coming to terms with the past’ began to include the ‘second German dictatorship’ in the form of the GDR. In the 1990s, GDR remembrance projects mushroomed, mainly centring on the border and the activities of the ‘Stasi’. It is tempting to view these remembrance debates in a Left versus Right, or anti‐fascist versus anti‐totalitarian, analytical framework. A case study of the Berlin Wall Memorial site and ‘Documentation Center’ in Bernauer Strasse shows that remembrance of the Wall indeed has high potential for instrumentalisation in political‐ideological conflicts. However, several other factors have also contributed to the development of this remembrance of the Wall, and its improvised form and content. Apart from an added East–West dimension, there are: the sheer number and diversity of participants and interested parties, the institutional fragmentation of competencies and responsibilities, a ‘psychology of resentment’, the scarcity of resources, the controversial aesthetics of remembrance, accidental factors such as post‐unification property legislation and the opposing roles of two church parishes, shifting political agendas and, finally, the demands of the tourism industry.
Anxiety and the Archive: Constructing Value and Identity in the Pop‐Lit Novels Soloalbum and Generation Golf
Authors: Pye Gillian
Published: March 2006

Recently the many angry responses in the German press provoked by examples of so‐called pop literature have testified to the challenge this writing presents to traditional conceptions of the cultural archive. By toying with the notions of transience and durability, the pop‐lit pose raises questions about the boundary between rubbish and the perennially valuable. Moreover, by focusing on the now as it becomes history –‘gerade, eben, jetzt’– it questions the relationship of literature to the past, reflecting a post‐memory generation's obsession with archiving the present. Florian Ilies's Generation Golf (2000) and Benjamin von Stuckrad Barre's Soloalbum (1998) are high‐profile examples of mainstream German pop literature of the late nineties. Critical commentators have tended to write off these texts as trash without assessing them on their own terms. This discussion considers Ilies's and von Stuckrad‐Barre's texts as provocative impulses which challenge literature to deal with the impact of consumer culture on the construction of identity.
The Jammed Democracy: Bolivia's Troubled Political Learning Process
Authors: Salman Ton
Published: March 2006

The fact that even the moderate and broadly respected president Carlos Mesa was forced to step down in Bolivia in June 2005 suggests that the country's crisis goes beyond a conflict on specific policies. A longstanding practice of excluding large sectors of the population from all real influence in politics, despite the existence of formal democracy, has produced a crisis of belief in democracy, affecting both governing bodies and the party system. President Mesa was unable to reverse the generalised distrust of politics. This distrust, combined with persisting political stalemate, is currently tending towards societal disintegration, which makes the recovery of genuine democratic practices even more difficult.
El Alto, Ciudad Rebelde: Organisational Bases for Revolt
Authors: Lazar Sian
Published: March 2006

The events of October 2003 reveal something about how Bolivian democracy works under ‘normal’ circumstances because they built upon well‐established patterns of political behaviour whereby corporate groupings have become used to direct negotiations with the government. It was the breakdown of these patterns that forced the resignation of the President in 2003. I detail here the collective identities that provide the foundations for mobilisational power in Bolivia and examine the organisation behind the uprising in El Alto and its roots in quotidian experiences of collective mobilisation. I conclude with a consideration of the relationship between social mobilisation, democracy and politics in contemporary Bolivia.
Praying for Government: Peasant Disengagement from the Bolivian State*
Authors: Goudsmit Into A.
Published: March 2006

This article explores peasants' experiences of the state in the Toracari valley, Northern Potosí, suggesting that such an analysis may shed light on the absence of political unrest among substantial segments of Bolivia's indigenous population. So long as Toracari peasants imagine the fetish of a benevolent (national) Government that one cannot fight, they associate local public offices with the small‐scale mestizo landlords who hold these positions, rather than with Government. Recognition of this distinction should caution scholars against employing the concept of ‘State’ as a system that includes all levels of government and state officials. Use of that concept may obstruct serious research into the power of local elites such as the landlords.
Stones on the Road: The Politics of Participation and the Generation of Crisis in Bolivia
Authors: Mcneish John‐Andrew
Published: March 2006

This paper demonstrates that recent protests in Bolivia must be linked to the failure of efforts to improve democratic participation in the country. It argues that such failures can be traced to a history of prejudices in national development and society and persistent biases and contradictions within international development policy and institutions. Despite these obstacles, the paper concludes that ideas for appropriate development and realistic alternatives for change to government and democracy are visible in recent critical development thinking and amongst the different social and cultural groupings involved in the demonstrations.
Todos somos indígenas: Towards a New Language of National Political Identity
Authors: Canessa Andrew
Published: March 2006

One of the most striking aspects of ‘Red October’ was the central role played by indigenous groups and their leaders who were largely able to set the agenda of protest. This paper explores how the concept of indigeneity as a mobilising concept has moved from the periphery of the political arena to centre stage. Two indigenous leaders played significant roles: whereas the political rhetoric of Felipe Quispe is exclusionary and particularistic, Evo Morales's rhetoric is inclusive and broad. Indigenous identity as articulated by indigenous leaders is contrasted to the identities expressed by rural people and raises the question of how indigeneity is defined and by whom.
Asserting State Authority through Environmental Monitoring: Venezuela in the Post‐Gómez Era, 1935–1945
Authors: Kozloff Nikolas
Published: March 2006

In Venezuela, the task of consolidating the central state fell to Presidents Eleazar López Contreras (1936–1941) and Isaías Medina Angarita (1941–1945). This meant asserting fiscal control over the oil companies operating in the state of Zulia as well as appeasing restive working and middle classes. The government sought to exert power over both by moving towards a system of environmental monitoring of the petroleum industry. López Contreras was reluctant to apply real political pressure to the companies and accordingly environmental monitoring tended to be lax. Medina Angarita personally visited zulianos and made fiery speeches designed to assuage Venezuelan pride. What is more, he allowed the press to denounce the companies for their environmental abuse. Through his environmental monitoring efforts, Medina demonstrated to the companies that the state was a force to be reckoned with.
Bacterial endotoxin as inhibitor of the enzymatic activity of human thrombin
Authors: Bucki Robert , Pastore Jennifer J.
Published: March 2006

Abstract:  Endotoxemia caused by bacterial lipopolysaccharides (LPS) deleteriously affects many aspects of hemostasis. Much of this effect is well characterized as being secondary to the LPS‐mediated inflammatory response, but direct effects of LPS on coagulation factors may also contribute to disregulation of the hemostatic process. Spectrophotometric assays were used to investigate the effects of LPS from different bacteria on thrombin and plasmin activities. We found that enzymatic activity of purified thrombin, but not plasmin, decreases in the presence of endotoxin. LPS‐mediated inhibition of thrombin activity can be reversed by plasma gelsolin and recombinant endotoxin‐neutralizing protein. Preincubation of thrombin with LPS before platelet activation results in inhibition of aggregation and secretion. Additionally, a decrease of elastic shear moduli of fibrin gels was observed when their formation was induced with thrombin preincubated with LPS or when LPS was present in fibrinogen solutions during fibrin gel formation. When added to platelet‐rich plasma, after activation with collagen, LPS‐inhibited thrombin activity. LPS‐mediated inhibition of thrombin activity may contribute to the hemostasis dysfunctions observed during endotoxemia.
Bortezomib: an effective agent in extramedullary disease in multiple myeloma
Authors: Laura Rosiñol , Cibeira Ma Teresa , Uriburu Carla , Yantorno Sebastián , Salamero Olga , Bladé Joan , Montserrat Emili
Published: March 2006

Abstract:  Bortezomib is a potent and selective proteasome inhibitor recently introduced in the treatment of multiple myeloma (MM). This drug produces significant responses in about one‐third of patients with relapsed/refractory disease. We first recognized the lack of efficacy of thalidomide in soft‐tissue plasmacytomas. There is little information on the effect of bortezomib on extramedullary myeloma. Four of 23 patients treated with bortezomib at our institution had extramedullary involvement at the time of relapse. In three of these patients large soft‐tissue plasmacytomas disappeared. This indicates that bortezomib may be useful in clinical situations of extramedullary disease in which other agents, such as thalidomide, may not be effective.