bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular biological study on speciation and phylogeny of the order Entomobryomorpha (Collembola: Hexapoda)
Authors: PARK Kyung‐Hwa
Published: September 2009

In order to elucidate the phylogenetic relationship among groups of the order Entomobryomorpha (Collembola), the sequences on theITS 1toITS 2fragments of the rRNA gene were analyzed in 11 species of three families. In order to avoid the potential risks and inconsistencies of a single method or data set, the phylogenetic reconstructions were based on three different approaches: methods of maximum parsimony, maximum likelihood and neighbor joining. The inferred phylogenies supported monophyly of the order Entomobryomorpha. The relationships between families were different, but the orders of branching within each family were the same. Entomobryidae and Isotomidae were paraphyletic, whereas Tomoceridae was monophyletic. Tomoceridae was subdivided into two branches; the molecular analysis provided results distinctive enough to separate the two genera by the high bootstrap value. On the other hand, two different populations of putativeHomidia koreanaappeared to be different species, although their chaetotaxy is identical. A wide coverage of characters, including not only morphological characters but also genetic data such as allozymes and DNA sequences, will give a more accurate picture of the classification and phylogeny of the studied group.
An evaluation of the SPIFE® 3000 semi‐automated gel electrophoresis system for the identification of hemoglobin variants and comparison of relative electrophoretic mobilities with manual gel electrophoresis methods
Authors: HOYER J. D. , MARKLEY K. M. , SAVEDRA M. E. , KUBIK K. S. , SCHEIDT R. M.
Published: September 2009

Laboratory identification of hemoglobin (Hb) variants can involve multiple techniques. The use of semi‐automated instruments that perform gel electrophoresis and staining, such as the SPIFE® 3000 electrophoresis system, can greatly reduce the labor required for these commonly used techniques. We performed a comparison of the method involved in SPIFE® 3000 system with those of manual gel electrophoresis. A total of 22 540 samples were analyzed using the SPIFE® 3000, and compared with mobilities on cellulose acetate and citrate agar gels using standard manual methods. The results were compared using relative electrophoretic mobilities (REM). Of the 191 Hb variants identified, only 13 had REM that differed from manual electrophoresis when analyzed using the SPIFE® 3000 system. One variant (Hb O‐Indonesia) showed different mobility on both acid and alkaline gels, two (Hb E, Hb Sunshine Seth) on alkaline gel only, and 10 (Hbs N‐Baltimore, N‐Seattle, O‐Arab, Shelby, Summer Hill, Tak, Hasharon, M‐Iwate, Q‐Iran, and Setif) on acid gels only. The SPIFE® 3000 semi‐automated electrophoresis system produces similar results when compared with those of standard manual electrophoresis methods.
Finding Common Ground: How Advocacy Coalitions Succeed in Protecting Environmental Flows1
Authors: Smith Mark P.
Published: September 2009

Abstract:  This paper reviews several recent case studies in which states or countries have strengthened their protection of environmental flows to explore the key policy, stakeholder, and scientific elements that contributed to these advances in water management. A conceptual framework is developed to describe the actions of interest groups and individuals, how environmental flow issues get onto the formal agenda of decision makers, the events and conditions which precipitate this attention, the role of science and scientific uncertainty, and how interactions and dialog among individuals and groups with different interests lead to changes in state and national statutes. In general, the review found that changing policies is a result of actions of informed groups of interested parties using science and information to inform both the public and decision makers about the need for action and about the specific action needed. In almost all cases, environmental flow issues make it onto the formal agenda of institutions through one or more precipitating events, often legal challenges that call into question the existing legal framework for water management. Significantly, in almost all cases the engagement between advocacy coalitions with different and often opposing views results in reframing the issues to provide a common approach or solution upon which the competing coalitions can agree.
A Metamodeling Technique for Variable Geometry Trusses Design via Equivalent Parametric Macroelements
Authors: Aguirrebeitia Josu , Angulo Carlos , Macareno Luis M. , Avilés Rafael
Published: September 2009

A metamodeling methodology is applied to reduce the large computational cost required in the design of variable geometry trusses (VGTs). Using this methodology, submodels of finite elements within a complete FE model of the VGT are substituted by groups of fewer elements called equivalent parametric macroelements (EPMs). The EPM optimum parameters are obtained using equivalence criteria based on elastic energy and inertial properties. The optimization process is performed using nonlinear least square minimization and genetic algorithms.
Human Immune Proteome in Experimental Colonization with
Staphylococcus aureus
Authors: Holtfreter Silva , Nguyen Thi Thu Hoai , Wertheim Heiman , Steil Leif , Kusch Harald , Truong Quoc Phong , Engelmann Susanne , Hecker Michael , Völker Uwe , van Belkum Alex , Bröker Barbara M.
Published: September 2009

More than 20% of adults are persistently colonized with
Staphylococcus aureus.
When hospitalized, these carriers have increased risks of infection with their own strains. However, a recent study demonstrated a lower incidence of bacteremia-related death among carriers than among noncarriers, raising the question whether the adaptive immune system plays a protective role. In fact,
S. aureus
carriers mount a highly specific neutralizing antibody response against superantigens of their colonizing strains. We now used 2-dimensional immunoblotting to investigate the profiles of antibodies from healthy individuals against
S. aureus
extracellular proteins. Moreover, we tested whether symptom-free experimental colonization of these individuals with an
S. aureus
strain of low virulence, 8325-4, is sufficient to induce an antibody response. Sera obtained before and 4 weeks after colonization were screened for immunoglobulin G (IgG) antibody binding to extracellular staphylococcal proteins. At baseline, most volunteers harbored IgG directed against conserved virulence factors, including alpha-hemolysin (Hla), beta-hemolysin (Hlb), phospholipase C (Plc), staphylococcal serine protease (SspA), and cysteine protease (SspB). However, the variability of spot patterns and intensities was striking and could be important in case of infection. Experimental nasal colonization with
S. aureus
8325-4 did not elicit new antibodies or boost the humoral response. Thus, the high antibody prevalence in humans is likely not induced by short-term nasal colonization, and presumably minor infections are required to trigger anti-
S. aureus
antibody responses.
Epidemiology and Clinical Presentations of Human Coronavirus NL63 Infections in Hong Kong Children
Authors: Leung Ting Fan , Li Chung Yi , Lam Wai Yip , Wong Gary W. K. , Cheuk Edmund , Ip Margaret , Ng Pak Cheung , Chan Paul K. S.
Published: September 2009

Human coronavirus NL63 (HCoV-NL63) has been found in children presenting with respiratory tract infections (RTIs). However, the epidemiology and clinical course of this newly identified virus have not been fully elucidated. This study investigated the epidemiology, seasonality, and clinical features of HCoV-NL63 in Hong Kong children. This study consisted of two cohorts of children hospitalized in a university-affiliated teaching hospital. In the 12-month retrospective part of the study, reverse transcription-PCR was used to detect HCoVs in nasopharyngeal aspirates (NPAs). Positive samples were sequenced to confirm the identity of the virus and to determine its phylogenetic relationship with the HCoV-NL63 strains found elsewhere. The second part covered a subsequent 12-month period in which patients were prospectively recruited. Altogether, 1,981 and 1,001 NPA specimens were studied in 2005-2006 and 2006-2007, respectively. Seventy-four (2.5%) HCoV isolates were identified and consisted of 17 (0.6%) HCoV-NL63 isolates, 37 (1.2%) HCoV-OC43 isolates, 14 (0.5%) HCoV-HKU1 isolates, and 6 (0.2%) HCoV-229E isolates. HCoV-NL63 infection was more common in 2006-2007 than 2005-2006 (1.2% and 0.3%, respectively;
P
= 0.006). From 2005 to 2007, the peak season for HCoV-NL63 infection was in September-October, which was earlier than the peak for HCoV-OC43 infections (December-January). HCoV-NL63-infected patients were younger and more likely to have croup, febrile convulsions, and acute gastroenteritis. The majority of local HCoV-NL63 isolates were phylogenetically closely related to those found in Belgium and The Netherlands. In conclusion, HCoV-NL63 is an important yet uncommon virus among our hospitalized children with acute RTIs.
Chronic Hepatitis, Hepatic Dysplasia, Fibrosis, and Biliary Hyperplasia in Hamsters Naturally Infected with a Novel Helicobacter Classified in the
H. bilis
Cluster
Authors: Fox J. G. , Shen Z. , Muthupalani S. , Rogers A. R. , Kirchain S. M. , Dewhirst F. E.
Published: September 2009

We recently described helicobacter-associated progressive, proliferative, and dysplastic typhlocolitis in aging (18- to 24-month-old) Syrian hamsters. Other pathogens associated with typhlocolitis in hamsters,
Clostridium difficile
,
Lawsonia intracellularis
, and
Giardia
spp., were not indentified. The presence of
Helicobacter
genus-specific DNA was noted by PCR in cecal and paraffin-embedded liver samples from aged hamsters by the use of
Helicobacter
-specific PCR primers. By 16S rRNA analysis, the
Helicobacter
sp. isolated from the liver tissue was identical to the cecal isolates from hamsters. The six hamster 16S rRNA sequences form a genotypic cluster most closely related to
Helicobacter
sp.
Flexispira
taxon 8, part of the
Helicobacter bilis
/
H. cinaedi
group. Livers from aged helicobacter-infected hamsters showed various stages of predominantly portocentric and, to a lesser extent, perivenular fibrosis. Within nodules, there was cellular atypia consistent with nodular dysplasia. The livers also exhibited a range of chronic active portal/interface and lobular inflammation, with significant portal hepatitis being present. The inflammation was composed of a mixture of lymphocytes, neutrophils, and macrophages, indicative of its chronic-active nature in these aged hamsters infected with
Helicobacter
spp. The isolation of novel
Helicobacter
spp., their identification by PCR from the diseased livers of aged hamsters, and their taxonomic classification as belonging to the
Helicobacter bilis
cluster strengthen the argument that
H. bilis
and closely related
Helicobacter
spp. play an etiological role in hepatobiliary disease in both animals and humans.
Mycobacterium tuberculosis
Strains with Highly Discordant Rifampin Susceptibility Test Results
Authors: Van Deun A. , Barrera L. , Bastian I. , Fattorini L. , Hoffmann H. , Kam K. M. , Rigouts L. , Rüsch-Gerdes S. , Wright A.
Published: September 2009

The objectives of this study were to investigate the origin of highly discordant rifampin (rifampicin) (RMP) drug susceptibility test results obtained for
Mycobacterium tuberculosis
strains during proficiency testing. Nine Supra-National Tuberculosis Reference Laboratories tested the RMP susceptibilities of 19 selected
M. tuberculosis
strains, using standard culture-based methods. The strains were classified as definitely resistant (R) (
n
= 6) or susceptible (S) (
n
= 2) or probably resistant (PR) (
n
= 8) or susceptible (PS) (
n
= 3) based on
rpoB
mutations and treatment outcome. All methods yielded a susceptible result for the two S and three PS strains lacking an
rpoB
mutation and a resistant result for one R strain with a Ser531Leu mutation and one PR strain with a double mutation. Although the remaining 12 R and PR strains had
rpoB
mutations (four Asp516Tyr, three Leu511Pro, two Leu533Pro, one each His526Leu/Ser, and one Ile572Phe), they were all susceptible by the radiometric Bactec 460TB or Bactec 960 MGIT methods. In contrast, only one was susceptible by the proportion method on Löwenstein-Jensen medium and two on Middlebrook 7H10 agar. Low-level but probably clinically relevant RMP resistance linked to specific
rpoB
mutations is easily missed by standard growth-based methods, particularly the automated broth-based systems. Further studies are required to confirm these findings, to determine the frequency of these low-level-resistant isolates, and to identify technical improvements that may identify such strains.
Molecular and Phenotypic Evaluation of
Lichtheimia corymbifera
(Formerly
Absidia corymbifera
) Complex Isolates Associated with Human Mucormycosis: Rehabilitation of
L. ramosa
Authors: Garcia-Hermoso Dea , Hoinard Damien , Gantier Jean-Charles , Grenouillet Frédéric , Dromer Françoise , Dannaoui Eric
Published: September 2009

Thirty-eight isolates (including 28 isolates from patients) morphologically identified as
Lichtheimia corymbifera
(formerly
Absidia corymbifera
) were studied by sequence analysis (analysis of the internal transcribed spacer [ITS] region of the ribosomal DNA, the D1-D2 region of 28S, and a portion of the elongation factor 1α [EF-1α] gene). Phenotypic characteristics, including morphology, antifungal susceptibility, and carbohydrate assimilation, were also determined. Analysis of the three loci uncovered two well-delimited clades. The maximum sequence similarity values between isolates from both clades were 66, 95, and 93% for the ITS, 28S, and EF-1α loci, respectively, with differences in the lengths of the ITS sequences being detected (763 to 770 bp for isolates of clade 1 versus 841 to 865 bp for isolates of clade 2). Morphologically, the shapes and the sizes of the sporangiospores were significantly different among the isolates from both clades. On the basis of the molecular and morphological data, we considered isolates of clade 2 to belong to a different species named
Lichtheimia ramosa
because reference strains CBS 269.65 and CBS 270.65 (which initially belonged to
Absidia ramosa
) clustered within this clade. As neotype
A. corymbifera
strain CBS 429.75 belongs to clade 1, the name
L. corymbifera
was conserved for clade 1 isolates. Of note, the amphotericin B MICs were significantly lower for
L. ramosa
than for
L. corymbifera
(
P
< 0.005) but were always ≤0.5 μg/ml for both species. Among the isolates tested, the assimilation of melezitose was positive for 67% of the
L. ramosa
isolates and negative for all
L. corymbifera
isolates. In conclusion, this study reveals that two
Lichtheimia
species are commonly associated with mucormycosis in humans.
Persistence of Transgene Expression Influences CD8+T-Cell Expansion and Maintenance following Immunization with Recombinant Adenovirus
Authors: Finn Jonathan D. , Bassett Jennifer , Millar James B. , Grinshtein Natalie , Yang Teng Chih , Parsons Robin , Evelegh Carole , Wan Yonghong , Parks Robin J. , Bramson Jonathan L.
Published: September 2009

Previous studies determined that the CD8+T-cell response elicited by recombinant adenovirus exhibited a protracted contraction phase that was associated with long-term presentation of antigen. To gain further insight into this process, a doxycycline-regulated adenovirus was constructed to enable controlled extinction of transgene expression in vivo. We investigated the impact of premature termination of transgene expression at various time points (day 3 to day 60) following immunization. When transgene expression was terminated before the maximum response had been attained, overall expansion was attenuated, yielding a small memory population. When transgene expression was terminated between day 13 and day 30, the memory population was not sustained, demonstrating that the early memory population was antigen dependent. Extinction of transgene expression at day 60 had no obvious impact on memory maintenance, indicating that maintenance of the memory population may ultimately become independent of transgene expression. Premature termination of antigen expression had significant but modest effects on the phenotype and cytokine profile of the memory population. These results offer new insights into the mechanisms of memory CD8+T-cell maintenance following immunization with a recombinant adenovirus.
The Nedd4-Type Rsp5p Ubiquitin Ligase Inhibits Tombusvirus Replication by Regulating Degradation of the p92 Replication Protein and Decreasing the Activity of the Tombusvirus Replicase
Authors: Barajas Daniel , Li Zhenghe , Nagy Peter D.
Published: September 2009

Recent in vitro proteomics screens revealed that many host proteins could interact with the replication proteins of
Tomato bushy stunt virus
(TBSV), which is a small, plus-stranded RNA virus (Z. Li, D. Barajas, T. Panavas, D. A. Herbst, and P. D. Nagy, J. Virol. 82:6911-6926, 2008). To further our understanding of the roles of host factors in TBSV replication, we have tested the effect of Rsp5p, which is a member of the Nedd4 family of E3 ubiquitin ligases. The full-length Rsp5p, via its WW domain, is shown to interact with p33 and the central portion of p92
pol
replication proteins. We find that overexpression of Rsp5p inhibits TBSV replication in
Saccharomyces cerevisiae
yeast, while downregulation of Rsp5p leads to increased TBSV accumulation. The inhibition is caused by Rsp5p-guided degradation of p92
pol
, while the negative effect on the p33 level is less pronounced. Interestingly, recombinant Rsp5p also inhibits TBSV RNA replication in a cell-free replication assay, likely due to its ability to bind to p33 and p92
pol
. We show that the WW domain of Rsp5p, which is involved in protein interactions, is responsible for inhibition of TBSV replication, whereas the HECT domain, involved in protein ubiquitination, is not necessary for Rsp5p-mediated inhibition of viral replication. Overall, our data suggest that direct binding between Rsp5p and p92
pol
reduces the stability of p92
pol
, with consequent inhibition of TBSV replicase activity.
The Crystal Structure of PF-8, the DNA Polymerase Accessory Subunit from Kaposi's Sarcoma-Associated Herpesvirus
Authors: Baltz Jennifer L. , Filman David J. , Ciustea Mihai , Silverman Janice Elaine Y. , Lautenschlager Catherine L. , Coen Donald M. , Ricciardi Robert P. , Hogle James M.
Published: September 2009

Kaposi's sarcoma-associated herpesvirus is an emerging pathogen whose mechanism of replication is poorly understood. PF-8, the presumed processivity factor of Kaposi's sarcoma-associated herpesvirus DNA polymerase, acts in combination with the catalytic subunit, Pol-8, to synthesize viral DNA. We have solved the crystal structure of residues 1 to 304 of PF-8 at a resolution of 2.8 Å. This structure reveals that each monomer of PF-8 shares a fold common to processivity factors. Like human cytomegalovirus UL44, PF-8 forms a head-to-head dimer in the form of a C clamp, with its concave face containing a number of basic residues that are predicted to be important for DNA binding. However, there are several differences with related proteins, especially in loops that extend from each monomer into the center of the C clamp and in the loops that connect the two subdomains of each protein, which may be important for determining PF-8's mode of binding to DNA and to Pol-8. Using the crystal structures of PF-8, the herpes simplex virus catalytic subunit, and RB69 bacteriophage DNA polymerase in complex with DNA and initial experiments testing the effects of inhibition of PF-8-stimulated DNA synthesis by peptides derived from Pol-8, we suggest a model for how PF-8 might form a ternary complex with Pol-8 and DNA. The structure and the model suggest interesting similarities and differences in how PF-8 functions relative to structurally similar proteins.
A Proteomic Approach To Identify Candidate Substrates of Human Adenovirus E4orf6-E1B55K and Other Viral Cullin-Based E3 Ubiquitin Ligases
Authors: Dallaire Frédéric , Blanchette Paola , Branton Philip E.
Published: September 2009

It has been known for some time that the human adenovirus serotype 5 (Ad5) E4orf6 and E1B55K proteins work in concert to degrade p53 and to regulate selective export of late viral mRNAs during productive infection. Both of these functions rely on the formation by the Ad5 E4orf6 protein of a cullin 5-based E3 ubiquitin ligase complex containing elongins B and C. E1B55K is believed to function as the substrate recognition module for the complex and, in addition to p53, Mre11 and DNA ligase IV have also been identified as substrates. To discover additional substrates we have taken a proteomic approach by using two-dimensional difference gel electrophoresis to detect cellular proteins that decrease significantly in amount in p53-null H1299 human lung carcinoma cells after expression of E1B55K and E4orf6 using adenovirus vectors. Several species were detected and identified by mass spectroscopy, and for one of these, integrin α3, we went on in a parallel study to confirm it as a
bone fide
substrate of the complex (F. Dallaire et al., J. Virol. 83:5329-5338, 2009). Although the system has some limitations, it may still be of some general use in identifying candidate substrates of any viral cullin-based E3 ubiquitin ligase complex, and we suggest a series of criteria for substrate validation.
Ty3 Nuclear Entry Is Initiated by Viruslike Particle Docking on GLFG Nucleoporins
Authors: Beliakova-Bethell Nadejda , Terry Laura J. , Bilanchone Virginia , DaSilva Rhonda , Nagashima Kunio , Wente Susan R. , Sandmeyer Suzanne
Published: September 2009

Yeast retrotransposons form intracellular particles within which replication occurs. Because fungal nuclear membranes do not break down during mitosis, similar to retroviruses infecting nondividing cells, the cDNA produced must be translocated through nuclear pore complexes. The
Saccharomyces cerevisiae
long terminal repeat retrotransposon Ty3 assembles its Gag3 and Gag3-Pol3 precursor polyproteins into viruslike particles in association with perinuclear P-body foci. These perinuclear clusters of Ty3 viruslike particles localized to sites of clustered nuclear pore complexes (NPCs) in a
nup120
Δ mutant, indicating that Ty3 particles and NPCs interact physically. The NPC channels are lined with nucleoporins (Nups) with extended FG (Phe-Gly) motif repeat domains, further classified as FG, FxFG, or GLFG repeat types. These domains mediate partitioning of proteins between the cytoplasm and the nucleus. Here we have systematically examined the requirements for FG repeat domains in Ty3 nuclear transport. The GLFG domains interacted in vitro with virus-like particle Gag3, and this interaction was disrupted by mutations in the amino-terminal domain of Gag3, which is predicted to lie on the external surface of the particles. Accordingly, Ty3 transposition was decreased in strains with the GLFG repeats deleted. The spacer-nucleocapsid domain of Gag3, which is predicted to be internal to the particle, interacted with GLFG repeats and nucleocapsid localized to the nucleus. We conclude that Ty3 particle docking on nuclear pores is facilitated by interactions between Gag3 and GLFG Nups and that nuclear entry of the preintegration complex is further promoted by nuclear localization signals within the nucleocapsid and integrase.
A Novel Picornavirus Associated with Gastroenteritis
Authors: Li Linlin , Victoria Joseph , Kapoor Amit , Blinkova Olga , Wang Chunlin , Babrzadeh Farbod , Mason Carl J. , Pandey Prativa , Triki Hinda , Bahri Olfa , Oderinde Bamidele Soji , Baba Marycelin Mandu , Bukbuk David Nadeba , Besser John M. , Bartkus Joanne M. , Delwart Eric L.
Published: September 2009

A novel picornavirus genome was sequenced, showing 42.6%, 35.2%, and 44.6% of deduced amino acid identities corresponding to the P1, P2, and P3 regions, respectively, of the Aichi virus. Divergent strains of this new virus, which we named salivirus, were detected in 18 stool samples from Nigeria, Tunisia, Nepal, and the United States. A statistical association was seen between virus shedding and unexplained cases of gastroenteritis in Nepal (
P
= 0.0056). Viruses with approximately 90% nucleotide similarity, named klassevirus, were also recently reported in three cases of unexplained diarrhea from the United States and Australia and in sewage from Spain, reflecting a global distribution and supporting a pathogenic role for this new group of picornaviruses.
The Minimum Replication Origin of Merkel Cell Polyomavirus Has a Unique Large T-Antigen Loading Architecture and Requires Small T-Antigen Expression for Optimal Replication
Authors: Kwun Hyun Jin , Guastafierro Anna , Shuda Masahiro , Meinke Gretchen , Bohm Andrew , Moore Patrick S. , Chang Yuan
Published: September 2009

Merkel cell polyomavirus (MCV) is a recently discovered human polyomavirus causing the majority of human Merkel cell carcinomas. We mapped a 71-bp minimal MCV replication core origin sufficient for initiating eukaryotic DNA replication in the presence of wild-type MCV large T protein (LT). The origin includes a poly(T)-rich tract and eight variably oriented, GAGGC-like pentanucleotide sequences (PS) that serve as LT recognition sites. Mutation analysis shows that only four of the eight PS are required for origin replication. A single point mutation in one origin PS from a naturally occurring, tumor-derived virus reduces LT assembly on the origin and eliminates viral DNA replication. Tumor-derived LT having mutations truncating either the origin-binding domain or the helicase domain also prevent LT-origin assembly. Optimal MCV replication requires coexpression of MCV small T protein (sT), together with LT. An intact DnaJ domain on the LT is required for replication but is dispensable on the sT. In contrast, PP2A targeting by sT is required for enhanced replication. The MCV origin provides a novel model for eukaryotic replication from a defined DNA element and illustrates the selective pressure within tumors to abrogate independent MCV replication.
Phosphorylation of the Nuclear Form of Varicella-Zoster Virus Immediate-Early Protein 63 by Casein Kinase II at Serine 186
Authors: Mueller Niklaus H. , Graf Laurie L. , Orlicky David , Gilden Don , Cohrs Randall J.
Published: September 2009

Varicella-zoster virus (VZV) open reading frame (ORF) 63 is abundantly transcribed in latently infected human ganglia and encodes a 278-amino-acid protein, IE63, with immediate-early kinetics. IE63 is expressed in the cytoplasm of neurons during VZV latency and in both the cytoplasm and the nucleus during productive infection; however, the mechanism(s) involved in IE63 nuclear import and retention has remained unclear. We constructed and identified a recombinant monoclonal antibody to detect a posttranslationally modified form of IE63. Analysis of a series of IE63 truncation and substitution mutants showed that amino acids 186 to 195 are required for antibody binding. Synthetic peptides corresponding to this region identified IE63 S186 as a target for casein kinase II phosphorylation. In addition, acidic charges supplied by E194 and E195 were required for antibody binding. Immunofluorescence analysis of VZV-infected MeWo cells using the recombinant monoclonal antibody detected IE63 exclusively in the nuclei of infected cells, indicating that casein kinase II phosphorylation of S186 occurs in the nucleus and possibly identifying an initial molecular event operative in VZV reactivation.
Multifaceted Sequence-Dependent and -Independent Roles for Reovirus FAST Protein Cytoplasmic Tails in Fusion Pore Formation and Syncytiogenesis
Authors: Barry Christopher , Duncan Roy
Published: September 2009

Fusogenic reoviruses utilize the FAST proteins, a novel family of nonstructural viral membrane fusion proteins, to induce cell-cell fusion and syncytium formation. Unlike the paradigmatic enveloped virus fusion proteins, the FAST proteins position the majority of their mass within and internal to the membrane in which they reside, resulting in extended C-terminal cytoplasmic tails (CTs). Using tail truncations, we demonstrate that the last 8 residues of the 36-residue CT of the avian reovirus p10 FAST protein and the last 20 residues of the 68-residue CT of the reptilian reovirus p14 FAST protein enhance, but are not required for, pore expansion and syncytium formation. Further truncations indicate that the membrane-distal 12 residues of the p10 and 47 residues of the p14 CTs are essential for pore formation and that a residual tail of 21 to 24 residues that includes a conserved, membrane-proximal polybasic region present in all FAST proteins is insufficient to maintain FAST protein fusion activity. Unexpectedly, a reextension of the tail-truncated, nonfusogenic p10 and p14 constructs with scrambled versions of the deleted sequences restored pore formation and syncytiogenesis, while reextensions with heterologous sequences partially restored pore formation but failed to rescue syncytiogenesis. The membrane-distal regions of the FAST protein CTs therefore exert multiple effects on the membrane fusion reaction, serving in both sequence-dependent and sequence-independent manners as positive effectors of pore formation, pore expansion, and syncytiogenesis.
Transcriptional Analysis of the Adeno-Associated Virus Integration Site
Authors: Dutheil Nathalie , Henckaerts Els , Kohlbrenner Erik , Linden R. Michael
Published: September 2009

The nonpathogenic human adeno-associated virus type 2 (AAV-2) has adopted a unique mechanism to site-specifically integrate its genome into the humanMBS85gene, which is embedded inAAVS1on chromosome 19. The fact that AAV has evolved to integrate into this ubiquitously transcribed region and that the chromosomal motifs required for integration are located a few nucleotides upstream of the translation initiation start codon ofMBS85suggests that the transcriptional activity ofMBS85might influence site-specific integration and thus might be involved in the evolution of this mechanism. In order to begin addressing this question, we initiated the characterization of the humanMBS85promoter region and compared its transcriptional activity to that of the AAV-2 p5 promoter. Our results clearly indicate thatAAVS1is defined by a complex transcriptional environment and that theMBS85promoter shares key regulatory elements with the viral p5 promoter. Furthermore, we provide evidence for bidirectionalMBS85promoter activity and demonstrate that the minimal motifs required for AAV site-specific integration are present in the 5′ untranslated region of the gene and play a posttranscriptional role in the regulation ofMBS85expression. These findings should provide a framework to further elucidate the complex interactions between the virus and its cellular host in this unique pathway to latency.
Hemodynamic instability during treatment of intracranial dural arteriovenous fistula and carotid cavernous fistula with Onyx: preliminary results and anesthesia considerations
Authors: Amiridze N , Darwish R
Published: September 2009

Background and aim

9 patients with 10 arteriovenous fistulas were treated with Onyx at our institution over a period of 19 months: 4 direct and indirect carotid–cavernous fistulas (CCFs) and 6 dural arteriovenous fistula (DAVFs). Complete occlusion was achieved with no recurrences or permanent complications in our small series. We report hemodynamic instability, including severe bradycardia and asystole, during embolization of DAVF and CCF with Onyx in several patients. These changes were reversible with interruption of Onyx injection and administration of atropine. No recurrence of symptoms after atropine administration was noted as Onyx embolization continued during the same session or during postprocedural 24 h monitoring. No adverse clinical consequences were noted.

Results

Bradycardia was observed in 4 cases, with a brief asystole in 2 of these patients during transarterial and transvenous Onyx delivery at cavernous sinus and orbital levels. Based on our observation, hemodynamic instability was demonstrated during Onyx injection into the vessels that were in close proximity to the trigeminal nerve or its branches, especially in low-flow/low-volume compartment and may represent a direct effect of dimethyl sulfoxide/Onyx on the trigeminal nerve, resulting in vagal response from trigeminocardiac reflex.

Conclusion

Proposed measures to prevent this complication include pretreatment with atropine or prophylactic placement of transvenous pacemakers in patients with underlying heart block and patients with contraindications to atropine use.
DynaCT evaluation of in-stent restenosis following Wingspan stenting of intracranial stenosis
Authors: Moskowitz S I , Kelly M E , Haynes J , Fiorella D
Published: September 2009

Objective and importance

To describe the use of DynaCT angiographic imaging for the evaluation of Wingspan in-stent restenosis (ISR).

Methods

Two patients were treated with Wingspan stenting and percutaneous transluminal angioplasty (Patient 1 had treatment of a severe stenosis of the right middle cerebral artery and patient 2 had severe stenosis of the left intracranial internal carotid artery. Both patients developed ISR and were evaluated with high resolution DynaCT angiographic imaging.

Results

DynaCT demonstrated circumferential soft tissue density material distributed within the stent as the cause of the stenosis visualized with conventional angiography.

Conclusions

These findings support the hypothesis that ISR is caused by neointimal proliferation, rather than vascular re-coil with stent collapse.
Reconstruction of the right anterior circulation with the Pipeline embolization device to achieve treatment of a progressively symptomatic, large carotid aneurysm
Authors: Fiorella D , Albuquerque F , Gonzalez F , McDougall C G , Nelson P K
Published: September 2009

Introduction

We present the use of the Pipeline embolization device (PED) to achieve reconstruction of the right anterior circulation in a patient with a dolichoectatic internal carotid artery (ICA) and middle cerebral artery (MCA) and an associated symptomatic, large, carotid-ophthalmic segment aneurysm.

Clinical presentation

A 36-year-old man presented with progressive right eye vision loss followed by sudden severe headache. Subsequent neuroimaging revealed a large right carotid-ophthalmic segment aneurysm and diffuse ectasia of the supraclinoid ICA and proximal MCA. A coil embolization of the aneurysm was performed without stent support. Over the next year, the patient experienced increasing headache and progressive bitemporal vision loss. Serial MRI showed progressive coil compaction and recanalization of the aneurysm.

Treatment

The right anterior circulation was reconstructed with a total of six PEDs that extended from the distal M1 segment of the MCA proximally into the distal cavernous segment of the ICA. Follow-up angiography at 1 and 4 months demonstrated progressive complete occlusion of the aneurysm and a reorganization of blood flow to the anterior cerebral and anterior choroidal arteries. MRI and radiographic imaging provided evidence of progressive contraction of the intra-aneurysmal thrombus. The patient's headaches resolved and serial visual field examinations have demonstrated gradual improvement after treatment.

Conclusion

Extensive cerebrovascular reconstructions that are not possible using commercially available endovascular devices can be achieved with Pipeline. The safety, efficacy and long term implications of such reconstructions are currently being defined.
NUMERICAL ANALYSIS OF THE GENERALIZED MAXWELL EQUATIONS (WITH AN ELLIPTIC CORRECTION) FOR CHARGED PARTICLE SIMULATIONS
Authors: CIARLET PATRICK , LABRUNIE SIMON
Published: September 2009

When computing numerical solutions to the Vlasov–Maxwell equations, the source terms in Maxwell's equations usually fail to satisfy the continuity equation. Since this condition is required for the well-posedness of Maxwell's equations, it is necessary to introduce generalized Maxwell's equations which remain well-posed when there are errors in the sources. These approaches, which involve a hyperbolic, a parabolic and an elliptic correction, have been recently analyzed mathematically. The goal of this paper is to carry out the numerical analysis for several variants of Maxwell's equations with an elliptic correction.
INFLUENCE OF SURFACE TENSION AND BOTTOM TOPOGRAPHY ON INTERNAL WAVES
Authors: ANH CUNG THE
Published: September 2009

Following the global strategy introduced recently by Bona, Lannes and Saut in Ref. 7, we derive here in a systematic way, and for a large class of scaling regimes, asymptotic models for the propagation of internal waves at the interface between two layers of immiscrible fluids of different densities, under the rigid lid assumption, the presence of surface tension and with uneven bottoms. The full (Euler) model for this situation is reduced to a system of evolution equations posed spatially on ℝd, d = 1, 2, which involve two nonlocal operators. The different asymptotic models are obtained by expanding the nonlocal operators and the surface tension term with respect to suitable small parameters that depend variously on the amplitude, wavelengths and depth ratio of the two layers. Furthermore, the consistency of these asymptotic systems with the full Euler equations is established.
SOCIALISM AND SOCIAL DIMENSION OF WORK — EMPLOYMENT POLICIES ON DISABLED GROUPS IN CHINA
Authors: CHAU RUBY C. M.
Published: September 2009

This paper is about employment policies on disabled people in China. It argues that the Chinese government has constantly emphasised the social dimension of work. Employment has therefore been regarded as an important means for disabled people to integrate into the society and to contribute to public interests. Various measures have been introduced in legislation and in policies on education and employment to enhance job opportunities for disabled people in the last two decades. However, the effectiveness of these measures is limited. In face of the keen competition in the expansion of the private market, many enterprises and welfare services do not have sufficient capacity or enthusiasm to promote employment for disabled groups. The paper argues it is important for the government to provide more support and incentives to enterprises, services, disabled people and their family in order to realise the social dimension of work among the disabled groups.
REVISITING THE FATHER-SON CONFLICTS IN CHINESE FAMILIES FROM A CULTURAL LENS: IMPLICATIONS FOR FAMILY-CENTERED PRACTICE IN CHINESE SOCIETIES
Authors: CHAN SIMON T. M.
Published: September 2009

A Chinese proverb reads "Wu Chou Bu Cheng Fu Zi"—無仇不成父子 (hatred bonds the father and his son). This well describes many of the father-son struggles found in clinical cases in Chinese families where the son rejects the father, who tries his best to get along with the son. The explanation for this phenomenon can be found in the field of family therapy (Bowen, 1976, 1985; Minuchin, 1996, 1993), which states that the son is being triangulated in the marital conflict and allied with the mother. However, there is no comprehensive account of the issue of father-son conflict and its inconsistency with classical Chinese concepts such as filial piety. This gap in the research can be addressed with interviews of Chinese families in a clinical context, in which the concept of family justice (大義滅親) can be deduced. This article not only unearths the concept of family justice in Chinese families, which hitherto has been rather hidden (Hall,1990), but also prepares that way to continue the research practice of conducting family interview on Chinese families. Through such conceptual discussions, classical concepts in Chinese culture can be recalled, and strategies for indigenous Chinese family therapy can be cultivated.
An environmental sensor, TRPV4 is a novel regulator of intracellular Ca2+ in human synoviocytes
Authors: Itoh Yuka , Hatano Noriyuki , Hayashi Hidetoshi , Onozaki Kikuo , Miyazawa Keiji , Muraki Katsuhiko
Published: September 2009

The activation of a vanilloid type 4 transient receptor potential channel (TRPV4) has an obligatory role in regulation of intracellular Ca2+ (Ca2+i) in several types of cells including vascular and sensory organs. In this study, we provide evidence that TRPV4 is a functional regulator of Ca2+i in human synoviocytes. Although significant expression of TRPV4 in synoviocytes from patients with (RA) and without (CTR) rheumatoid arthritis was detected at mRNA and protein level, those in the human fibroblast-like synoviocyte line MH7A were rather lower. Consistently, the selective TRPV4 agonist 4α-phorbol 12,13-didecanoate (4αPDD) effectively elevated Ca2+i in the RA and CTR cells, which was abolished by the removal of external Ca2+. Moreover, the elevation was inhibited by ruthenium red, a blocker of TRPVs. In MH7A cells transfected with human TRPV4 (MH7A-V4), 4αPDD elevated the Ca2+i in a similar manner to those in the RA and CTR cells. Electrophysiological analysis also revealed that 4αPDD activated nonselective cationic currents in RA cells. Application of 227 mosM solution to the RA and MH7A-V4 cells elevated their Ca2+i, but this does not occur when it was applied to MH7A cells. Treatment of RA but not MH7A cells with 4αPDD for 24 h reduced their production of IL-8. These results suggest that an environmental sensor, TRPV4, is a novel regulator of intracellular Ca2+ in human synoviocytes.
Intramuscular VEGF repairs the failing heart: role of host-derived growth factors and mobilization of progenitor cells
Authors: Zisa David , Shabbir Arsalan , Mastri Michalis , Suzuki Gen , Lee Techung
Published: September 2009

Skeletal muscle produces a myriad of mitogenic factors possessing cardiovascular regulatory effects that can be explored for cardiac repair. Given the reported findings that VEGF may modulate muscle regeneration, we investigated the therapeutic effects of chronic injections of low doses of human recombinant VEGF-A165 (0.1–1 μg/kg) into the dystrophic hamstring muscle in a hereditary hamster model of heart failure and muscular dystrophy. In vitro, VEGF stimulated proliferation, migration, and growth factor production of cultured C2C12 skeletal myocytes. VEGF also induced production of HGF, IGF2, and VEGF by skeletal muscle. Analysis of skeletal muscle revealed an increase in myocyte nuclear [531 ± 12 VEGF 1 μg/kg vs. 364 ± 19 for saline (number/mm2) saline] and capillary [591 ± 80 VEGF 1 μg/kg vs. 342 ± 21 for saline (number/mm2)] densities. Skeletal muscle analysis revealed an increase in Ki67+ nuclei in the VEGF 1 μg/kg group compared with saline. In addition, VEGF mobilized c-kit+, CD31+, and CXCR4+ progenitor cells. Mobilization of progenitor cells was consistent with higher SDF-1 concentrations found in hamstring, plasma, and heart in the VEGF group. Echocardiogram analysis demonstrated improvement in left ventricular ejection fraction (0.60 ± 0.02 VEGF 1 μg/kg vs. 0.45 ± 0.01 mm for saline) and an attenuation in ventricular dilation [5.59 ± 0.12 VEGF 1 μg/kg vs. 6.03 ± 0.09 for saline (mm)] 5 wk after initiating therapy. Hearts exhibited higher cardiomyocyte nuclear [845 ± 22 VEGF 1 μg/kg vs. 519 ± 40 for saline (number/mm2)] and capillary [2,159 ± 119 VEGF 1 μg/kg vs. 1,590 ± 66 for saline (number/mm2)] densities. Myocardial analysis revealed ∼2.5 fold increase in Ki67+ cells and ∼2.8-fold increase in c-kit+ cells in the VEGF group, which provides evidence for cardiomyocyte regeneration and progenitor cell expansion. This study provides novel evidence of a salutary effect of VEGF in the cardiomyopathic hamster via induction of myogenic growth factor production by skeletal muscle and mobilization of progenitor cells, which resulted in attenuation of cardiomyopathy and repair of the heart.
Mice lacking the G protein γ3-subunit show resistance to opioids and diet induced obesity
Authors: Schwindinger William F. , Borrell Brandon M. , Waldman Lora C. , Robishaw Janet D.
Published: September 2009

Contributing to the obesity epidemic, there is increasing evidence that overconsumption of high-fat foods may be analogous to drug addiction in that the palatability of these foods is associated with activation of specific reward pathways in the brain. With this perspective, we report that mice lacking the G protein γ3-subunit ( Gng3−/−mice) show resistance to high-fat diet-induced weight gain over the course of a 12-wk study. Compared with Gng3+/+controls, female Gng3−/−mice exhibit a 40% reduction in weight gain and a 53% decrease in fat pad mass, whereas male Gng3−/−mice display an 18% reduction in weight gain and no significant decrease in fat pad mass. The basis for the lowered weight gain is related to reduced food consumption for female and male Gng3−/−mice of 13% and 14%, respectively. Female Gng3−/−mice also show a lesser preference for high-fat chow than their female Gng3+/+littermates, suggesting an attenuated effect on a reward pathway associated with overconsumption of fat. One possible candidate is the μ-opioid receptor ( Oprm1) signaling cascade. Supporting a defect in this signaling pathway, Gng3−/−mice show marked reductions in both acute and chronic morphine responsiveness, as well as increases in endogenous opioid mRNA levels in reward-related regions of the brain. Taken together, these data suggest that the decreased weight gain of Gng3−/−mice may be related to a reduced rewarding effect of the high-fat diet resulting from a defect in Oprm1 signaling and loss of the G protein γ3-subunit.
Peripheral osmotic stimulation inhibits the brain's innate immune response to microdialysis of acidic perfusion fluid adjacent to supraoptic nucleus
Authors: Summy-Long Joan Y. , Hu Sanmei
Published: September 2009

During the brain's innate immune response microglia, astroglia and ependymal cells resolve/repair damaged tissue and control infection. Released interleukin-1β (IL-1β) reaching cerebroventricles stimulates circumventricular organs (CVOs; subfornical organ, SFO; organum vasculosum lamina terminalis, OVLT), the median preoptic nucleus (MePO), and magnocellular and parvocellular neurons in the supraoptic (SON) and paraventricular (PVN) nuclei. Hypertonic saline (HS) also activates these osmosensory CVOs and neuroendocrine systems, but, in contrast to IL-1β, inhibits the peripheral immune response. To examine whether the brain's innate immune response is attenuated by osmotic stimulation, sterile acidic perfusion fluid was microdialyzed (2 μl/min) in the SON area of conscious rats for 6 h with sterile HS (1.5 M NaCl) injected subcutaneously (15 ml/kg) at 5 h. Immunohistochemistry identified cytokine sources (IL-1β+; OX-42+microglia) and targets (IL-1R+; inducible cyclooxygenase, COX-2+; c-Fos+) near the probe, in CVOs, MePO, ependymal cells, periventricular hypothalamus, SON, and PVN. Inserting the probe stimulated magnocellular neurons (c-Fos+; SON; PVN) via the MePO (c-Fos+), a response enhanced by HS. Microdialysis activated microglia (OX-42+; amoeboid/hypertrophied; IL-1β+) in the adjacent SON and bilaterally in perivascular areas of the PVN, periventricular hypothalamus and ependyma, coincident with c-Fos expression in ependymal cells and COX-2 in the vasculature. These microglial responses were attenuated by HS, coincident with activating parvocellular and magnocellular neuroendocrine systems and elevating circulating IL-1β, oxytocin, and vasopressin. Acidosis-induced cellular injury from microdialysis activated the brain's innate immune response by a mechanism inhibited by peripheral osmotic stimulation.