bluuebunny/crossref_metadata_2025 huggingface dataset:
The wound repair response controls outcome to cutaneous leishmaniasis
Authors: Sakthianandeswaren Anuratha , Elso Colleen M. , Simpson Ken , Curtis Joan M. , Kumar Beena , Speed Terence P. , Handman Emanuela , Foote Simon J.
Published: October 2005

Chronic microbial infections are associated with fibrotic and inflammatory reactions known as granulomas showing similarities to wound-healing and tissue repair processes. We have previously mapped three leishmaniasis susceptibility loci, designated
lmr1
, -
2
, and -
3
, which exert their effect independently of T cell immune responses. Here, we show that the wound repair response is critically important for the rapid cure in murine cutaneous leishmaniasis caused by
Leishmania major
. Mice congenic for leishmaniasis resistance loci, which cured their lesions more rapidly than their susceptible parents, also expressed differentially genes involved in tissue repair, laid down more ordered collagen fibers, and healed punch biopsy wounds more rapidly. Fibroblast monolayers from these mice repaired
in vitro
wounds faster, and this process was accelerated by supernatants from infected macrophages. Because these effects are independent of T cell-mediated immunity, we conclude that the rate of wound healing is likely to be an important component of innate immunity involved in resistance to cutaneous leishmaniasis.
Perspectives on the geographic stability and mobility of people in cities
Authors: Hanson Susan
Published: October 2005

A class of questions in the human environment sciences focuses on the relationship between individual or household behavior and local geographic context. Central to these questions is the nature of people's geographic mobility as well as the duration of their locational stability at varying spatial and temporal scales. The problem for researchers is that the processes of mobility/stability are temporally and spatially dynamic and therefore difficult to measure. Whereas time and space are continuous, analysts must select levels of aggregation for both length of time in place and spatial scale of place that fit with the problem in question. Previous work has emphasized mobility and suppressed stability as an analytic category. I focus here on stability and show how analyzing individuals' stability requires also analyzing their mobility. Through an empirical example centered on the relationship between entrepreneurship and place, I demonstrate how a spotlight on stability illuminates a resolution to the measurement problem by highlighting the interdependence between the time and space dimensions of stability/mobility.
Effect of protein structure on mitochondrial import
Authors: Wilcox Alexander J. , Choy Jason , Bustamante Carlos , Matouschek Andreas
Published: October 2005

Most proteins that are to be imported into the mitochondrial matrix are synthesized as precursors, each composed of an N-terminal targeting sequence followed by a mature domain. Precursors are recognized through their targeting sequences by receptors at the mitochondrial surface and are then threaded through import channels into the matrix. Both the targeting sequence and the mature domain contribute to the efficiency with which proteins are imported into mitochondria. Precursors must be in an unfolded conformation during translocation. Mitochondria can unfold some proteins by changing their unfolding pathways. The effectiveness of this unfolding mechanism depends on the local structure of the mature domain adjacent to the targeting sequence. This local structure determines the extent to which the unfolding pathway can be changed and, therefore, the unfolding rate increased. Atomic force microscopy studies find that the local structures of proteins near their N and C termini also influence their resistance to mechanical unfolding. Thus, protein unfolding during import resembles mechanical unfolding, and the specificity of import is determined by the resistance of the mature domain to unfolding as well as by the properties of the targeting sequence.
Human microRNAs target a functionally distinct population of genes with AT-rich 3′ UTRs
Authors: Robins Harlan , Press William H.
Published: October 2005

While investigating microRNA targets, we have found that human genes divide into two roughly equal populations, based on the fraction of A plus T bases in their 3′ UTRs. Using the Gene Ontology database, we find significant functional differences between the two gene populations, with AT-rich genes implicated in transcription and translation processes, and GC-rich genes implicated in signal transduction and posttranslational protein modification. Better understanding of the background distribution of nucleotides in 3′ UTRs may allow improved prediction of microRNA-targeted genes in humans. We predict at least 1,200 KnownGene transcripts to be regulated by microRNAs. The large majority of these microRNA targets are in the AT-rich 3′ UTR population. However, notwithstanding this preference for AT-rich targets, microRNA targets are found preferentially to be regulatory genes themselves, including both transcription factors and posttranslational modifiers. These results suggest that some processes involving mRNA, of which microRNA regulation may be just one, require AT-richness of 3′ UTRs for functionality. A relationship, not simply one-to-one, between these 3′ UTR populations and large-scale genomic isochores is described.
Oscillatory activity in the infant brain reflects object maintenance
Authors: Kaufman Jordy , Csibra Gergely , Johnson Mark H.
Published: October 2005

The apparent failure of infants to understand “object permanence” by reaching for hidden objects is perhaps the most striking and debated phenomenon in cognitive development. Of particular interest is the extent to which infants perceive and remember objects in a similar way to that of adults. Here we report two findings that clarify infant object processing. The first is that 6-mo-old infants are sensitive to visual cues to occlusion, particularly gradual deletion. The second finding is that oscillatory electroencephalogram activity recorded over right temporal channels is involved in object maintenance. This effect occurs only after disappearance in a manner consistent with occlusion and the object's continued existence.
Structure of the KvAP voltage-dependent K
+
channel and its dependence on the lipid membrane
Authors: Lee Seok-Yong , Lee Alice , Chen Jiayun , MacKinnon Roderick
Published: October 2005

Voltage-dependent ion channels gate open in response to changes in cell membrane voltage. This form of gating permits the propagation of action potentials. We present two structures of the voltage-dependent K
+
channel KvAP, in complex with monoclonal Fv fragments (3.9 Å) and without antibody fragments (8 Å). We also studied KvAP with disulfide cross-bridges in lipid membranes. Analyzing these data in the context of the crystal structure of Kv1.2 and EPR data on KvAP we reach the following conclusions: (
i
) KvAP is similar in structure to Kv1.2 with a very modest difference in the orientation of its voltage sensor; (
ii
) mAb fragments are not the source of non-native conformations of KvAP in crystal structures; (
iii
) because KvAP contains separate loosely adherent domains, a lipid membrane is required to maintain their correct relative orientations, and (
iv
) the model of KvAP is consistent with the proposal of voltage sensing through the movement of an arginine-containing helix-turn-helix element at the protein-lipid interface.
Isoform Diversity of Giant Proteins in Relation to Passive and Active Contractile Properties of Rabbit Skeletal Muscles
Authors: Prado Lucas G. , Makarenko Irina , Andresen Christian , Krüger Martina , Opitz Christiane A. , Linke Wolfgang A.
Published: October 2005

The active and passive contractile performance of skeletal muscle fibers largely depends on the myosin heavy chain (MHC) isoform and the stiffness of the titin spring, respectively. Open questions concern the relationship between titin-based stiffness and active contractile parameters, and titin's importance for total passive muscle stiffness. Here, a large set of adult rabbit muscles (n = 37) was studied for titin size diversity, passive mechanical properties, and possible correlations with the fiber/MHC composition. Titin isoform analyses showed sizes between ∼3300 and 3700 kD; 31 muscles contained a single isoform, six muscles coexpressed two isoforms, including the psoas, where individual fibers expressed similar isoform ratios of 30:70 (3.4:3.3 MD). Gel electrophoresis and Western blotting of two other giant muscle proteins, nebulin and obscurin, demonstrated muscle type–dependent size differences of ≤70 kD. Single fiber and single myofibril mechanics performed on a subset of muscles showed inverse relationships between titin size and titin-borne tension. Force measurements on muscle strips suggested that titin-based stiffness is not correlated with total passive stiffness, which is largely determined also by extramyofibrillar structures, particularly collagen. Some muscles have low titin-based stiffness but high total passive stiffness, whereas the opposite is true for other muscles. Plots of titin size versus percentage of fiber type or MHC isoform (I-IIB-IIA-IID) determined by myofibrillar ATPase staining and gel electrophoresis revealed modest correlations with the type I fiber and MHC-I proportions. No relationships were found with the proportions of the different type II fiber/MHC-II subtypes. Titin-based stiffness decreased with the slow fiber/MHC percentage, whereas neither extramyofibrillar nor total passive stiffness depended on the fiber/MHC composition. In conclusion, a low correlation exists between the active and passive mechanical properties of skeletal muscle fibers. Slow muscles usually express long titin(s), predominantly fast muscles can express either short or long titin(s), giving rise to low titin-based stiffness in slow muscles and highly variable stiffness in fast muscles. Titin contributes substantially to total passive stiffness, but this contribution varies greatly among muscles.
Improving disulfide connectivity prediction with sequential distance between oxidized cysteines
Authors: Tsai Chi-Hung , Chen Bo-Juen , Chan Chen-hsiung , Liu Hsuan-Liang , Kao Cheng-Yan
Published: October 2005

Summary: Predicting disulfide connectivity precisely helps towards the solution of protein structure prediction. In this study, a descriptor derived from the sequential distance between oxidized cysteines (denoted as DOC) is proposed. An approach using support vector machine (SVM) method based on weighted graph matching was further developed to predict the disulfide connectivity pattern in proteins. When DOC was applied, prediction accuracy of 63% for our SVM models could be achieved, which is significantly higher than those obtained from previous approaches. The results show that using the non-local descriptor DOC coupled with local sequence profiles significantly improves the prediction accuracy. These improvements demonstrate that DOC, with a proper scaling scheme, is an effective feature for the prediction of disulfide connectivity. The method developed in this work is available at the web server PreCys (prediction of cys–cys linkages of proteins).

Availability:  

Contact:  cykao@csie.ntu.edu.tw

Supplementary information: Supplementary data, detailed results, tables and information are available at
Fertilization‐Induced Changes in Growth Parameters and Antioxidant Activity of Medicinal Plants Used in Traditional Arab Medicine
Authors: Azaizeh Hassan , Ljubuncic Predrag , Portnaya Irina , Said Omar , Cogan Uri , Bomzon Arieh
Published: October 2005

In response to increased popularity and greater demand for medicinal plants, a number of conservation groups are recommending that wild medicinal plants be brought into cultivation systems. We collected four medicinal herbs Cichorium pumilum, Eryngium creticum, Pistacia palaestina and Teucrium polium used in traditional Arab medicine for greenhouse cultivation to assess the effects of different fertilization regimes on their growth and antioxidant activity. Wild seedlings were collected and fertilized with either 100% Hoagland solution, 50% Hoagland solution, 20% Hoagland solution or irrigated with tap water. Plant height was measured and the number of green leaves and branches counted weekly. Thereafter, the aboveground parts of plants were harvested for preparing a water‐soluble powder extracts of which antioxidant activity was measured by their ability to suppress the oxidation of β‐carotene. Of the fertilization regimes, we found either 20 or 50% Hoagland solution produced the most consistent response of the plant growth parameters. All powders prepared from the four wild growing plants inhibited oxidation of β‐carotene. Increasing the amount of fertilizer caused a significant concentration‐dependent increase in antioxidant activity of the cultivated T. polium compared with the wild type. In contrast, increasing the amount of fertilizer caused a significant concentration‐dependent reduction in the antioxidant activity of powders prepared from the cultivated E. creticum when compared with wild plants. Our results showed that cultivation success should not rely solely on parameters of growth but should incorporate assessment related to indices of therapeutic potential.
Surfactant protein D binds to human immunodeficiency virus (HIV) envelope protein gp120 and inhibits HIV replication
Authors: Meschi Joseph , Crouch Erika C. , Skolnik Paul , Yahya Khabirah , Holmskov Uffe , Leth-Larsen Rikke , Tornoe Ida , Tecle Tesfaldet , White Mitchell R. , Hartshorn Kevan L.
Published: October 2005

The envelope protein (gp120) of human immunodeficiency virus (HIV) contains highly conserved mannosylated oligosaccharides. These glycoconjugates contribute to resistance to antibody neutralization, and binding to cell surface lectins on macrophages and dendritic cells. Mannose-binding lectin (MBL) binds to gp120 and plays a role in defence against the virus. In this study it is demonstrated that surfactant protein D (SP-D) binds to gp120 and inhibits HIV infectivity at significantly lower concentrations than MBL. The binding of SP-D was mediated by its calcium-dependent carbohydrate-binding activity and was dependent on glycosylation of gp120. Native dodecameric SP-D bound to HIV gp120 more strongly than native trimeric SP-D. Since one common polymorphic form of SP-D is predominantly expressed as trimers and associated with lower blood levels, these individuals may have less effective innate defence against HIV. A chimeric protein containing the N-terminal and collagen domains of SP-D linked to the neck and carbohydrate-recognition domains of MBL (called SP-D/MBLneck+CRD) had greater ability to bind to gp120 and inhibit virus replication than either SP-D or MBL. The enhanced binding of SP-D/MBLneck+CRDwas dependent on assembly into higher molecular mass multimers (i.e. a trimeric form of the chimera did not bind to a greater extent than MBL). Hence, the enhanced binding of SP-D compared with MBL results from distinctive properties of its N-terminal and/or collagen domains. SP-D is present in lung and airway fluids, as well as in blood and various mucosal locations, and could, like MBL, play a role in restricting HIV transmission or replicationin vivo.
Characterization and mapping of monoclonal antibodies against the Sleeping disease virus, an aquatic alphavirus
Authors: Moriette Coralie , LeBerre Monique , Boscher Soasig Kerbart , Castric Jeannette , Brémont Michel
Published: October 2005

Sleeping disease virus (SDV) is an emerging pathogen in salmonid aquacultures, the impact of which is underestimated to date due to the lack of efficient diagnostic tools. To better characterize this new aquatic alphavirus and to make molecular tools available, a panel of monoclonal antibodies (mAbs) directed against SDV non-structural and structural proteins has been generated by immunizing mice with SDV-specific recombinant proteins overexpressed in Escherichia coli as antigens. So far, mAbs against nsP1, nsP3, E2 and E1 SDV proteins have been produced and their reactivity has been characterized by Western blot, radioimmunoprecipitation and indirect immunofluorescence assays. In addition, protein domains recognized by each mAb have been determined by immunofluorescence assay on truncated expressed SDV-derived proteins. Finally, one mAb directed against the E1 glycoprotein has been evaluated as a potential tool to be used in immunohistochemistry assay on experimentally infected trout.
Trypsin is associated with the rotavirus capsid and is activated by solubilization of outer capsid proteins
Authors: Benureau Yann , Huet Jean Claude , Charpilienne Annie , Poncet Didier , Cohen Jean
Published: October 2005

The rotavirus capsid is made up of three concentric protein layers. The outer layer, consisting of VP7 and VP4, is lost during virus entry into the host cell. Rotavirus field isolates can be adapted to high-titre growth in tissue culture by treatment with trypsin and by supplementing the culture medium with trypsin, which cleaves VP4 into two fragments, VP8* and VP5*. It is known that protease inhibitors reduce the replication of rotavirusin vitroandin vivoand also diminish disease symptoms in a mouse model. To clarify the molecular basis of these observations, a series of assays were conducted on purified rotavirus particles grown in the presence of trypsin. Results of HPLC and mass spectrometry followed by N-terminal sequencing showed that viral particles contain molecules of trypsin. When associated with triple-layer particles (TLPs), trypsin is inactive and not accessible to protease inhibitors, such as aprotinin. When the outer layer is solubilized by calcium-chelating agents, VP5*, VP8* and VP7 are released and the associated trypsin is activated, allowing cleavage of the viral capsid proteins, as well as other exogenous proteins. It is shown that addition of trypsin inhibitors significantly reduces synthesis of viral mRNA and viral proteins in cells and has a major inhibitory effect if present when virus enters the cell. These data indicate that incorporation of trypsin into rotavirus particles may enhance its infectivity.
Semliki Forest virus-derived virus-like particles: characterization of their production and transduction pathways
Authors: Diatta A. , Piver E. , Collin C. , Vaudin P. , Pagès J.-C.
Published: October 2005

A procedure for the mobilization of Semliki Forest virus (SFV)-derived replicons using virus-like particles (VLPs) has been recently proposed. VLPs were obtained from 293T cells co-expressing the vesicular stomatitis virus glycoprotein (VSV-G) and a modified SFV replicon. Advantages of SFV VLPs include improved safety with a lack of sequence homology between components and reducing the risk of recombination events that could lead to the formation of autonomous particles. Characterization of SFV VLPs reveals a discrepancy in their ability to infect cells reported to be permissive. Furthermore, it was noted that not all viral envelopes were able to promote VLP release equally from transfected cells. These observations encouraged the examination of the molecular mechanisms supporting the different steps of VLP assembly and transduction. The use of a VSV-G related pathway for VLP entry into target cells was demonstrated; it was also observed that an internal ribosome entry site may not be adapted to control transgene expression in all cells. Finally, the need for a membrane-binding domain to obtain a fully active SFV replication complex and VLP formation was documented.
Characterization of a highly glycosylated form of the human cytomegalovirus HLA class I homologue gpUL18
Authors: Griffin Cora , Wang Eddie C. Y. , McSharry Brian P. , Rickards Carole , Browne Helena , Wilkinson Gavin W. G. , Tomasec Peter
Published: October 2005

Human cytomegalovirus (HCMV) gpUL18 is a HLA class I (HLA-I) homologue with high affinity for the inhibitory receptor LIR-1/ILT2. The previously described 67 kDa form of gpUL18 is shown here to be sensitive to endoglycosidase-H (EndoH). A novel form of gpUL18 with a molecular mass of ∼160 kDa and resistance to EndoH was identified in cells infected with HCMV strain AD169 or the low passage HCMV isolates Merlin and Toledo. The 67 kDa EndoH-sensitive gpUL18 glycoform was detected earlier in a productive infection (from 24 h post-infection) than the slower-migrating EndoH-resistant glycoform (from 72 h post-infection). Deletion of the US2–US11 region from the HCMV genome was associated with a substantial up-regulation of endogenous HLA-I in infected cells, but had no obvious effect on the gpUL18 expression pattern. Vaccinia virus and adenovirus vectors were used to further analyse gpUL18 expression. Depending on the delivery vector system, differences in the electrophoretic motility of the EndoH-resistant >105 kDa form of gpUL18, but not the EndoH-sensitive 67 kDa form, were observed; post-translational modification of the higher molecular mass glycoform appears to be influenced by active virus infection and vector delivery. The EndoH-sensitive 67 kDa gpUL18 had a rapid turnover, while the maturation to the EndoH-resistant >105 kDa form was relatively slow and inefficient. However, synthesis of the EndoH-resistant >105 kDa form was enhanced with elevated levels ofβ2-microglobulin. When expressed by using an adenovirus vector, both the EndoH-sensitive 67 kDa and the EndoH-resistant >105 kDa gpUL18 forms could be detected on the cell surface.
Evidence for interaction between the 2a polymerase protein and the 3a movement protein of Cucumber mosaic virus
Authors: Hwang Min Sook , Kim Sang Hyon , Lee Jeong Hyun , Bae Jung Myung , Paek Kyung Hee , Park Young In
Published: October 2005

The genome of Cucumber mosaic virus consists of three single-stranded RNA molecules, RNAs 1, 2 and 3. RNAs 1 and 2 encode the 1a and 2a proteins, respectively, which are necessary for replication of the viral genome and have been implicated in movement of the viral RNAs in plants. The 3a movement protein (MP), encoded by RNA 3, is essential for transferring the RNA genomes from infected cells to adjacent cells across the plasmodesmata. Far-Western analysis demonstrated that bacterially expressed 2a polymerase protein directly interacted with the MP. Interaction was confirmed in a yeast two-hybrid assay, and co-immunoprecipitation analysis showed that the MP interacted only with the 2a polymerase protein. A yeast three-hybrid assay showed that the 1a–2a protein interaction relevant for replicase complex formation was not affected by the MP. Although the MP has no affinity for the 1a protein, it interacted indirectly with the 1a protein via the 2a polymerase protein. These results suggest that the replicase complex may be involved in movement through its interaction with the MP.
An in vitro model for the regulation of human cytomegalovirus latency and reactivation in dendritic cells by chromatin remodelling
Authors: Reeves M. B. , Lehner P. J. , Sissons J. G. P. , Sinclair J. H.
Published: October 2005

Human cytomegalovirus (HCMV) is a frequent cause of major disease following primary infection or reactivation from latency in immunocompromised patients. Infection of non-permissive mononuclear cells is used for analyses of HCMV latency in vitro. Using this approach, it is shown here that repression of lytic gene expression following experimental infection of CD34+ cells, a site of HCMV latency in vivo, correlates with recruitment of repressive chromatin around the major immediate-early promoter (MIEP). Furthermore, long-term culture of CD34+ cells results in carriage of viral genomes in which the MIEP remains associated with transcriptionally repressive chromatin. Finally, specific differentiation of long-term cultures of infected CD34+ cells to mature dendritic cells results in acetylation of histones bound to the MIEP, concomitant loss of heterochromatin protein 1 and the reactivation of HCMV. These data are consistent with ex vivo analyses of latency and may provide a model for further analyses of the mechanisms involved during latency and reactivation.
Signal peptide peptidase promotes the formation of hepatitis C virus non-enveloped particles and is captured on the viral membrane during assembly
Authors: Majeau Nathalie , Gagné Valérie , Bolduc Marilène , Leclerc Denis
Published: October 2005

The maturation of the core protein (C) ofHepatitis Cvirus(HCV) is controlled by the signal peptidase (sp) and signal peptide peptidase (spp) of the host. To date, it remains unknown whether spp cleavage influences viral infectivity and/or the assembly process. Here, evidence is provided that cleavage by spp is not required for assembly of nucleocapsid-like particles (NLPs) in yeast (Pichia pastoris). The immature NLPs (not processed by spp) show a density of 1·11 g ml−1on sucrose gradients and a diameter of 50 nm. Co-expression of human spp (hspp) with C generates the 21 kDa mature form of the protein and promotes the accumulation of non-enveloped particles. The amount of non-enveloped particles accumulating in the cell was correlated directly with the expression level of hspp. Furthermore, immunocapture studies showed that hspp was embedded in the membranes of enveloped particles. These results suggest that maturation of the C protein can occur after formation of the enveloped particles and that the abundance of hspp influences the types of particle accumulating in the cells.
Recombinant poxviruses as mucosal vaccine vectors
Authors: Gherardi M. Magdalena , Esteban Mariano
Published: October 2005

The majority of infections initiate their departure from a mucosal surface, such asHuman immunodeficiency virus(HIV), a sexually transmitted virus. Therefore, the induction of mucosal immunity is a high priority in the development of vaccines against mucosal pathogens. The selection of an appropriate antigen delivery system is necessary to induce an efficient mucosal immune response. Poxvirus vectors have been the most intensively studied live recombinant vector, and numerous studies have demonstrated their ability to induce mucosal immune responses against foreign expressed antigens. Previous studies have demonstrated that recombinants based on the attenuated modified vaccinia virus Ankara (MVA) vector were effective in inducing protective responses against different respiratory viruses, such as influenza and respiratory syncytial virus, following immunization via mucosal routes. Recent studies performed in the murine and macaque models have shown that recombinant MVA (rMVA) does not only stimulate HIV-specific immunity in the genital and rectal tracts following mucosal delivery, but can also control simian/human immunodeficiency viraemia and disease progression. In addition, a prime-boost vaccination approach against tuberculosis emphasized the importance of the intranasal rMVA antigen delivery to induce protective immunity againstMycobacterium tuberculosis. The aim of this review is to summarize the studies employing recombinant poxviruses, specifically rMVA as a mucosal delivery vector. The results demonstrate that rMVAs can activate specific immune responses at mucosal surfaces, and encourage further studies to characterize and improve the MVA mucosal immunogenicity of poxvirus vectors.
A novel recombinant of Hepatitis B
virus genotypes G and C isolated from a Thai patient with hepatocellular carcinoma
Authors: Suwannakarn Kamol , Tangkijvanich Pisit , Theamboonlers Apiradee , Abe Kenji , Poovorawan Yong
Published: October 2005

Genomic recombination between different genotypes of Hepatitis B
virus (HBV) resulting in hybrid strains has been increasingly documented. In this study, a novel recombinant of HBV genotypes G and C isolated from a Thai patient with hepatocellular carcinoma is reported. Based on phylogenetic analyses of the S, P and X genes and the entire genome, the HBV isolate clustered on a branch within genotype G, but clustered with genotype C on analysis of the C gene. Using the program simplot and bootscanning analysis, the recombination breakpoints were located at nt 1860 and 2460 of the precore/core region. The hallmarks of the original genotype G, including a 36 bp insertion in the core region and dual stop codons in the precore region, were not identified in this isolate. These data should encourage further investigations on the epidemiological and virological characteristics of HBV genotype G involved in recombination with other genotypes.
Vaccinia virus intracellular enveloped virions move to the cell periphery on microtubules in the absence of the A36R protein
Authors: Herrero-Martínez Esteban , Roberts Kim L. , Hollinshead Michael , Smith Geoffrey L.
Published: October 2005

Vaccinia virus (VACV) intracellular enveloped virus (IEV) particles are transported to the cell periphery on microtubules where they fuse with the plasma membrane to form cell-associated enveloped virus (CEV). Two IEV-specific proteins, F12L and A36R, are implicated in mediating transport of IEV. Without F12L, virus morphogenesis halts after formation of IEV, and CEV is not formed, whereas without A36R, IEV was reported not to be transported, yet CEV was formed, To address the roles of A36R and F12L in IEV transport, viruses with deletions of either F12L (vΔF12L) or A36R (vΔA36R) were labelled with enhanced green fluorescent protein (EGFP) fused to the core protein A5L, and used to follow CEV production with time. Without F12L, CEV production was inhibited by >99 %, whereas without A36R, CEV were produced at ∼60 % of wild-type levels at 24 h post-infection. Depolymerization of microtubules, but not actin, inhibited CEV formation in vΔA36R-infected cells. Moreover, vΔA36R IEV labelled with EGFP fused to the B5R protein co-localized with microtubules, showing that the A36R protein is not required for the interaction of IEV with microtubules. Time-lapse confocal microscopy confirmed that both wild-type and vΔA36R IEV moved in a stop–start manner at speeds consistent with microtubular movement, although the mean length of vΔA36R IEV movement was shorter. These data demonstrate that VACV IEV is transported to the cell surface using microtubules in the absence of A36R, and therefore IEV must attach to microtubule motors using at least one protein other than A36R.
IE1 and hr facilitate the localization of Bombyx mori nucleopolyhedrovirus ORF8 to specific nuclear sites
Authors: Kang WonKyung , Imai Noriko , Kawasaki Yu , Nagamine Toshihiro , Matsumoto Shogo
Published: October 2005

The Bombyx mori nucleopolyhedrovirus (BmNPV) ORF8 protein has previously been reported to colocalize with IE1 to specific nuclear sites during infection. Transient expression of green fluorescent protein (GFP)-fused ORF8 showed the protein to have cytoplasmic localization, but following BmNPV infection the protein formed foci, suggesting that ORF8 requires some other viral factor(s) for this. Therefore, interacting factors were looked for using the yeast two-hybrid system and IE1 was identified. We mapped the interacting region of ORF8 using a yeast two-hybrid assay. An N-terminal region (residues 1–110) containing a predicted coiled-coil domain interacted with IE1, while a truncated N-terminal region (residues 1–78) that lacks this domain did not. In addition, a protein with a complete deletion of the N-terminal region failed to interact with IE1. These results suggest that the ORF8 N-terminal region containing the coiled-coil domain is required for the interaction with IE1. Next, whether IE1 plays a role in ORF8 localization was investigated. In the presence of IE1, GFP-ORF8 localized to the nucleus. In addition, cotransfection with a plasmid expressing IE1 and a plasmid containing the hr3 element resulted in nuclear foci formation. A GFP-fused ORF8 mutant protein containing the coiled-coil domain, previously shown to interact with IE1, also formed nuclear foci in the presence of IE1 and hr3. However, ORF8 mutant proteins that did not interact with IE1 failed to form nuclear foci. In contrast to wild-type IE1, focus formation was not observed for an IE1 mutant protein that was deficient in hr binding. These results suggest that IE1 and hr facilitate the localization of BmNPV ORF8 to specific nuclear sites.
A comparison of haemagglutination, haemagglutination inhibition and PCR for the detection of psittacine beak and feather disease virus infection and a comparison of isolates obtained from loriids
Authors: Khalesi Bahman , Bonne Nicolai , Stewart Meredith , Sharp Margaret , Raidal Shane
Published: October 2005

Psittacine beak and feather disease (PBFD) is recognized as a threat for endangered psittacine birds in Australia, New Zealand and South Africa. Several diagnostic methods for the detection of beak and feather disease virus (BFDV) infection have been developed but there are few studies comparing the relative merits or sensitivity and specificity of each diagnostic test. In this report, the results of PCR, haemagglutination (HA) and haemagglutination inhibition (HI) testing of diagnostic samples collected from 679 samples from a range of psittacine bird species suspected of being infected with BFDV are summarized and compared. There was a strong agreement (kappa = 0·757;P<0·0001) between PCR and HA testing of feather samples and PCR-negative birds were 12·7 times more likely to have HI antibody than PCR-positive birds. False-positive HA results with titres up to 1 : 320 were identified in six feather samples that were PCR negative; the haemagglutination detected in these samples was not inhibited by anti-BFDV antisera and was removed by filtration through a 0·22 μm filter. Similarly, one false-negative PCR result was detected in a feather sample that had a high HA titre (>1 : 40 960) and four false-positive PCR results were detected in a batch of four feather samples. Of 143 birds that were feather PCR positive, only two had detectable HI antibody, and these birds were also feather HA negative, suggesting that they were developing immunity to recent infection. All birds with HI antibody were negative on feather HA testing. The assays confirmed BFDV infection in two endangered swift parrots (Lathamus discolor) and phylogenetic analysis of the sequence data generated from ORF V1 of these isolates provide further evidence of BFDV genotypes clustering in parallel with the Loriidae, Cacatuidae and Psittacidae.
Replication kinetics of Marek's disease vaccine virus in feathers and lymphoid tissues using PCR and virus isolation
Authors: Baigent Susan J. , Smith Lorraine P. , Currie Richard J. W. , Nair Venugopal K.
Published: October 2005

CVI988 (Rispens), an avirulent strain of Marek's disease virus, is the most widely used vaccine against Marek's disease. The kinetics of replication of CVI988 was examined in tissues of chickens vaccinated at either 1 day or 14 days of age and sampled regularly up to 28 days post-vaccination. Age at vaccination had no significant effect on the kinetics of CVI988 virus replication. During the cytolytic phase of infection (1–7 days), virus levels peaked in the spleen, bursa and thymus with very close correlation among these organs. Virus load in peripheral blood lagged behind and did not reach high levels. Significant numbers of virus genomes were detected in the feather tips only after 7 days, but subsequently rose to levels almost 103-fold greater than in the other tissues. This is the first accurate quantitative data for kinetics of CVI988 replication in a variety of tissues. There was good correlation between data from virus isolation and PCR, with real-time PCR being the preferred method for rapid, accurate and sensitive quantification of virus. Feathers were ideal for non-invasive sampling to detect and measure CVI988 in live chickens and, from 10 days onwards, virus load in feather tips was predictive of virus load in lymphoid tissues where immune responses will occur. The potential for real-time PCR analysis of feather samples for further investigation of the mechanism of vaccinal protection, and to assist optimization of vaccination regimes, is discussed.
Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon
Authors: Targett-Adams Paul , McLauchlan John
Published: October 2005

Dicistronic, subgenomic hepatitis C virus (HCV) replicons were constructed containing sequences from JFH1, a genotype 2a strain, that also incorporated the firefly luciferase gene under the control of the HCV internal ribosome entry site element. Luciferase activity in Huh-7 cell extracts containing in vitro-transcribed subgenomic JFH1 RNA was monitored over a 72 h period to examine early stages of HCV replication in the absence of any selective pressure. Enzyme activities produced by the replicon were almost 200-fold greater than those generated from corresponding genotype 1b replicons and correlated with an accumulation of NS5A protein and replicon RNA. Transient replication was sensitive to IFN treatment in a dose-dependent manner and, in addition to Huh-7 cells, the U2OS human osteosarcoma cell line supported efficient replication of the JFH1 replicon. Thus, this system based on JFH1 sequences offers improvements over prior genotype 1b replicons for quantitative measurement of viral RNA replication.
Traumatic Brain Injury and Hypopituitarism
Authors: Aimaretti Gianluca , Ghigo Ezio
Published: October 2005

Results of recent and ongoing studies have made it clear that brain injuries like Traumatic Brian Injury (TBI) pose substantial risk to pituitary function, perhaps even greater risk than previously believed. Patients with TBI should be screened both prospectively and retrospectively for isolated, multiple and even total pituitary deficits. It is well known that, patients with “classical” hypopituitarism (due to primary hypothalamic-pituitary pathologies) do benefit from hormonal replacement therapy. It has been suggested that patients with TBI-induced hypopituitarism may benefit with appropriate hormonal replacement receiving replacement therapy such as anti-diuretic hormone (ADH), glucocorticoid and thyroid hormones when needed. Gonadal and recombinant human Growth Hormone (rhGH) replacement therapy should also be introduced if there are deficiencies demonstrated and even reconfirmed in a second step. The signs and symptoms of post-TBI hypopituitarism may be masked by what has been assumed to be merely the post-traumatic syndrome. By increasing awareness among physicians of the risks of brain injuries–induced endocrinopathies and the need for appropriate endocrinological testing, it may be possible to improve the quality of life and enhance the rehabilitation prospects for these patients. In most instances, these patients are first seen and treated by trauma surgeons and neurosurgeons, and subsequently by rehabilitation physicians; they must be knowledgeable about the risks of hypopituitarism so that they can determine which patients are candidates for screening for hypopituitarism. In addition, endocrinologists and internists must be educated about TBI-induced hypopituitarism and encouraged to actively share their expertise with other physicians.
Chemokines in Renal Diseases
Authors: Segerer Stephan , Nelson Peter J.
Published: October 2005

The chemokines, members of a large family of chemotactic cytokines, act as directional cues for sorting inflammatory cell subsets to sites of inflammation or lymphoid microenvironments. In addition to their effects on migration, chemokines can also activate effector function in leukocytes and are involved in cell proliferation and angiogenesis. Therefore, it is not surprising that chemokines play important roles in a wide range of human diseases, including genetic immunodeficiencies, infections, autoimmune diseases, and malignant tumors. In this report, we have reviewed recent developments (since mid 2003) in chemokines in renal diseases. In animal models, chemokines are produced at the site of injury, leading to inflammatory cell recruitment. The therapeutic impact of the blockade of CCR1, CCR2, CCR4, CCR5, or the corresponding ligands has been further studied in various renal disease models. Recent studies on the role of the chemokine receptors in human diseases have demonstrated the expression of CXCR1, CXCR3, CCR2, and CCR5 on different subsets of inflammatory cells. The number of CCR5- and CXCR3-positive interstitial infiltrating cells (mainly T cells) correlates with renal function and proteinuria in glomerular diseases. Polymorphisms of chemokines and chemokine receptors are of impact on renal disease courses and allograft survival. Chemokine receptor blockade has approached clinical applications in nonrenal diseases and awaits the application in patients with kidney diseases.
The Diagnosis of Minimal Change Disease in Diabetic Nephropathy
Authors: Stokes M. Barry
Published: October 2005

Proteinuria is common in diabetic patients and usually reflects the presence of diabetic glomerulosclerosis. This paper reviews the differential diagnosis of proteinuria in diabetic patients and discusses the role of renal biopsy examination in identification and management of minimal change disease in this cohort. Identification of nondiabetic glomerular disease requires careful correlation of clinical history and renal biopsy findings and may have important implications for prognosis and therapy.
Clinical Holistic Medicine: Holistic Rehabilitation
Authors: Ventegodt Søren , Gringols Mark , Merrick Joav
Published: October 2005

Quality of life, health, and ability are often lost at the same time and most often in one decaying existential movement over 5 or 10 years. This “lost life” is mostly too slow to be felt as life threatening, but once awakened to reality, it provokes the deepest of fears in patients: the fear of death itself and destruction of our mere existence. The horrible experience of having “lost life”, often without even noticing how it happened, can be turned into a strong motivation for improvement. Personal development is about finding the life deeply hidden within in order to induce revitalization and rehabilitation. Rehabilitation is about philosophy of life with the integration of the repressed painful feelings and emotions from the past and the letting go of the associated negative beliefs and decisions. The holistic medical toolbox builds on existential theories (the quality of life theories, the life mission theory, the theory of character, the theory of talent, and the holistic process theory) and seems to have the power to rehabilitate the purpose of life, the character of the person, and fundamental existential dimensions of man: (1) love; (2) strength of mind, feelings, and body; and 3) joy, gender, and sexuality; allowing the person once again to express and realize his talents and full potential. The principles of rehabilitation are not very different from other healing, but the task is often more demanding for the holistic physician as the motivation and resources often are very low and the treatment can take many years.
Transient Hydroureteronephrosis Caused by a Foley's Catheter Tip in the Right Ureter
Authors: George Jacob , Tharion George
Published: October 2005

We report a case of unilateral hydronephrosis following urethral catheterization in a patient with T6 complete paraplegia at the Physical Medicine and Rehabilitation Department in a tertiary care teaching hospital, India. Diagnosis was established by an abdominal ultrasound. The misplaced catheter tip was withdrawn from the ureteric orifice and hydronephrosis was resolved. Foley's catheterization, a widely practiced clinical procedure, is not without its attendant risks of an inadvertent placement in the ureter leading to transient hydronephrosis. Inadequate drainage through a catheter should thus alert one to this potentially hazardous complication that can be diagnosed by an early ultrasound. This complication can be avoided by gently tugging on the catheter after inflating the catheter bulb.
Clinical Holistic Medicine: Whiplash, Fibromyalgia, and Chronic Fatigue
Authors: Ventegodt Søren , Gringols Mark , Merrick Joav
Published: October 2005

Holistic treatment of the highly complex, “new diseases” are often possible with the tools of consciousness-based medicine. The treatment is more complicated and the cure usually takes longer than for less-complex diseases. The problem with these patients is that they have less easily accessible resources than most patients, as they suffer from a combined socio-psycho-physical problem with depression, poor social standing, low confidence, and low self-esteem. Often, they have also already tried most of the specialist and alternative treatments on the market. To cure them, the most important thing is to coach them to improve their social life by changing their behavior to be of more value to others. Holding and processing must be especially careful and the contract with the patients must be extremely explicit in order to work on their personal development for 6—12 months. The new diseases can be cured with consciousness-based medicine if the patients are motivated and keep their appointments and agreements. Low responsibility, low personal energy, little joy of life, and limited insight into self and existence are some of the features of the new diseases that make them difficult to cure. The important thing is to keep a pace the patient can follow and give the patient a row of small successes and as few failures as possible. The new diseases are a challenge, a unique chance to improve communication, holding, and processing skills.
Interaction of Arsenic with Zinc and Organics in a Rice (Oryza sativaL.)–Cultivated Field in India
Authors: Das Dilip Kumar , Garai T. K. , Sarkar S. , Sur Pintu
Published: October 2005

A laboratory experiment on an Inceptisol with pH 7.6, organic carbon 6.8 g kg–1, and 0.5MNaHCO3extractable arsenic 0.4 mg kg–1was conducted to study the interaction effect of graded levels of arsenic (0, 5, and 10 mg kg–1) with zinc (0, 10, and 20 mg kg–1) and organics (0, 1, and 2% on soil weight basis) separately on the mobilization of arsenic in soils.The results show that the amount of 0.5MNaHCO3extractable arsenic at pH 8.5 increased with the progress of submergence up to 35 days. However, the increase in arsenic concentration was correlated with decreasing application of graded levels of Zn as zinc sulfate. The intensity of reduction varied with varying levels of Zn, being higher (0.73–2.72 mg kg–1) in the treatment where Zn was at 10 mg kg–1and lower (0.70–1.08 mg kg–1) with Zn at 20 mg kg–1application.The amount of arsenic content in the soil significantly decreased with the application of varying levels of organics. However, such depressive effect was found more pronounced with well-decomposed farm yard manure than that of vermicompost. The results of field experiments showed that the grain yield between continuous flooding (4.84 t ha–1) and intermittent flooding up to 40 days after transplanting then continuous flooding (4.83 t ha–1) with the application of ZnSO4at 25 kg ha–1did not vary. The lowest grain yield (3.65 t ha–1) was recorded in the treatment where intermittent flooding was maintained throughout the growth period without the application of Zn. The amount of arsenic content was, however, recorded much lower in the treatment where intermittent flooding throughout the growth period was maintained with ZnSO4.