bluuebunny/crossref_metadata_2025 huggingface dataset:
Comment on “On the motion of an ice hockey puck,” by K. Voyenli and E. Eriksen [Am. J. Phys. 53 (12), 1149–1153 (1985)]
Authors: Denny Mark
Published: May 2006

Voyenli and Eriksen [Am. J. Phys. 53, 1149–1153 (1985)] have derived some interesting and surprising results for a thin homogeneous disk (a hockey puck or a compact disk) and for a thin ring that is sliding and rotating over a flat but rough surface, assuming sliding friction with a constant coefficient. The properties that were observed and derived include the following: (a) When started with a pure translation, the disk will continue until it comes to rest in pure rectilinear translation without being set into rotation. (b) When started as a pure rotation around its center, the disk will continue to rotate while the center remains at rest until the disk stops. (c) In both cases the time it takes for the disk to come to rest increases with increasing values of the initial velocity. (d) When started with a combination of translation and rotation, the center will continue in a rectilinear motion until (e) the translation and rotation stop simultaneously, regardless of the initial velocity. The surprise is (e), which implies that there is a frictional coupling between the translation and rotation of the disk. This result was shown in Voyenli and Eriksen (1985) where it is established that for a thin ring of radius R, the final stage of the motion is rolling-like, meaning that the disk slides with an angular speed ω proportional to its linear speed v, so that w≡ωR=v.
Einstein, Perrin, and the reality of atoms: 1905 revisited
Authors: Newburgh Ronald , Peidle Joseph , Rueckner Wolfgang
Published: May 2006

We have repeated Perrin’s 1908 experiment for the determination of Avogadro’s number by determining the mean square displacement of small particles undergoing Brownian motion. Our apparatus differs from Perrin’s by the use of a CCD camera and is much less tedious to perform. We review Einstein’s 1905 analysis of Brownian motion and Langevin’s alternative derivation of the Einstein equation for the mean square displacement. We also show how Einstein’s thinking was a reflection of his belief in the validity of molecular-kinetic theory, a validity not universally recognized 100 years ago.
Type 1 Interferons and the Virus-Host Relationship: A Lesson in Détente
Authors: García-Sastre Adolfo , Biron Christine A.
Published: May 2006

The interface between an infectious agent and its host represents the ultimate battleground for survival: The microbe must secure a niche for replication, whereas the host must limit the pathogen's advance. Among the host's arsenal of antimicrobial factors, the type 1 interferons (IFNs) induce potent defense mechanisms against viruses and are key in the host-virus standoff. Viruses have evolved multiple tricks to avoid the immediate antiviral effects of IFNs and, in turn, hosts have adapted use of this innate cytokine system to galvanize multiple additional layers of immune defense. The plasticity that exists in these interactions provides us with a lesson in détente.
Aggresomes and Autophagy Generate Sites for Virus Replication
Authors: Wileman Thomas
Published: May 2006

The replication of many viruses is associated with specific intracellular compartments called virus factories or virioplasm. These are thought to provide a physical scaffold to concentrate viral components and thereby increase the efficiency of replication. The formation of virus replication sites often results in rearrangement of cellular membranes and reorganization of the cytoskeleton. Similar rearrangements are seen in cells in response to protein aggregation, where aggresomes and autophagosomes are produced to facilitate protein degradation. Here I review the evidence that some viruses induce aggresomes and autophagosomes to generate sites of replication.
Roles of Human Parainfluenza Virus Type 3 Bases 13 to 78 in Replication and Transcription: Identification of an Additional Replication Promoter Element and Evidence for Internal Transcription Initiation
Authors: Hoffman Michael A. , Thorson LeeAnne M. , Vickman John E. , Anderson Joseph S. , May Nathan A. , Schweitzer Michelle N.
Published: May 2006

The genomic promoter of human parainfluenza virus type 3 (HPIV3) contains multiple
cis
-elements controlling transcription and replication. Previous work showed that regions 1 to 12 and 79 to 96 were critical in promoting replication of an HPIV3 minireplicon, while the intergenic sequence and N gene start signal (IS/Ngs, bases 49 to 61) were important for transcription. Because these data were collected primarily using point mutations, not every base from position 1 to 96 was analyzed, and some important control elements may have been missed. To clarify the role of bases 13 to 78 in transcription and replication, a series of mutations were made which collectively scanned this entire region. Mutation of bases 13 to 28 resulted in markedly decreased HPIV3 minireplicon replication, indicating these bases constitute an additional
cis
-element involved in the synthesis of the HPIV3 antigenomic RNA. The position dependence of the IS/Ngs was also examined. Analysis of mutants in which the IS/Ngs was shifted 5′ or 3′ showed that this segment could be moved without significantly disrupting transcription initiation. Additional mutants which contained two successive IS/Ngs segments were created to test whether the polymerase accessed the gene start signal by proceeding along the template 3′ to 5′ or by binding internally at the gene start signal. Based on analysis of the double gene start mutants, we propose a model of internal transcription initiation in which the polymerase enters the template at approximately the location of the natural N gene start but then scans the template bidirectionally to find a gene start signal and initiate transcription.
Two Surface-Exposed Elements of the B30.2/SPRY Domain as Potency Determinants of N-Tropic Murine Leukemia Virus Restriction by Human TRIM5α
Authors: Perron Michel J. , Stremlau Matthew , Sodroski Joseph
Published: May 2006

Human TRIM5α (TRIM5α
hu
) potently restricts N-tropic (N-MLV), but not B-tropic, murine leukemia virus in a manner dependent upon residue 110 of the viral capsid. Rhesus monkey TRIM5α (TRIM5α
rh
) inhibits N-MLV only weakly. The study of human-monkey TRIM5α chimerae revealed that both the v1 and v3 variable regions of the B30.2/SPRY domain contain potency determinants for N-MLV restriction. These variable regions are predicted to be surface-exposed elements on one face of the B30.2 domain. Acidic residues in v3 complement basic residue 110 of the N-MLV capsid. The results support recognition of the retroviral capsid by the TRIM5α B30.2 domain.
Silencing of Hepatitis A Virus Infection by Small Interfering RNAs
Authors: Kusov Yuri , Kanda Tatsuo , Palmenberg Ann , Sgro Jean-Yves , Gauss-Müller Verena
Published: May 2006

Infection by hepatitis A virus (HAV) can cause acute hepatitis and, rarely, fulminant liver failure, in particular in patients chronically infected with hepatitis C virus. Based on our previous observation that small interfering RNAs (siRNAs) can silence translation and replication of the firefly luciferase-encoding HAV replicon, we now exploited this technology to demonstrate the effect of siRNAs on viral infection in Huh-7 cells. Freshly and persistently infected cells were transfected with siRNAs targeting various sites in the HAV nonstructural genes. Compared to a single application, consecutive siRNA transfections targeting multiple sequences in the viral genome resulted in a more efficient and sustained silencing effect than a single transfection. In most instances, multiple applications of a single siRNA led to the emergence of viral escape mutants with mutated target sites that rendered these genomes resistant to RNA interference (RNAi). Efficient and sustained suppression of the viral infectivity was achieved after consecutive applications of an siRNA targeting a computer-predicted hairpin structure. This siRNA holds promise as a therapeutic tool for severe courses of HAV infection. In addition, the results provide new insight into the structural bases for sequence-specific RNAi.
Furin Cleavage Potentiates the Membrane Fusion-Controlling Intersubunit Disulfide Bond Isomerization Activity of Leukemia Virus Env
Authors: Sjöberg Mathilda , Wallin Michael , Lindqvist Birgitta , Garoff Henrik
Published: May 2006

The membrane fusion protein of murine leukemia virus is a trimer of a disulfide-linked peripheral-transmembrane (SU-TM) subunit complex. The intersubunit disulfide bond is in SU linked to a disulfide bond isomerization motif, CXXC, with which the virus controls its fusion reaction (M. Wallin, M. Ekström, and H. Garoff, EMBO J. 23:54-65, 2004). Upon receptor binding the isomerase rearranges the intersubunit disulfide bond into a disulfide bond isomer within the motif. This facilitates SU dissociation and fusion activation in the TM subunit. In the present study we have asked whether furin cleavage of the Env precursor potentiates the isomerase to be triggered. To this end we accumulated the late form of the precursor, gp90, in the cell by incubation in the presence of a furin-inhibiting peptide. The isomerization was done by NP-40 incubation or by a heat pulse under alkylation-free conditions. The cells were lysed in the presence of alkylator, and the precursor was immunoprecipitated, gel isolated, deglycosylated, and subjected to complete trypsin digestion. Disulfide-linked peptide complexes were separated by sodium dodecyl sulfate-tricine-polyacrylamide gel electrophoresis under nonreducing conditions. This assay revealed the size of the characteristic major disulfide-linked peptide complex that differentiates the two isomers of the disulfide bond between Cys336 (or Cys339) and Cys563, i.e., the bond corresponding to the intersubunit disulfide bond. The analyses showed that the isomerase was five- to eightfold more resistant to triggering in the precursor than in the mature, cleaved form. This suggests that the isomerase becomes potentiated for triggering by a structural change in Env that is induced by furin cleavage in the cell.
Human Papillomavirus Type 16 E6 Activates NF-κB, Induces cIAP-2 Expression, and Protects against Apoptosis in a PDZ Binding Motif-Dependent Manner
Authors: James Michael A. , Lee John H. , Klingelhutz Aloysius J.
Published: May 2006

Infection with human papillomavirus (HPV) is a critical factor in the pathogenesis of most cervical cancers and some aerodigestive cancers. The HPV E6 oncoprotein from high-risk HPV types contributes to the immortalization and transformation of cells by multiple mechanisms, including degradation of p53, transcriptional activation of human telomerase reverse transcriptase (hTERT), and degradation of several proteins containing PDZ domains. The ability of E6 to bind PDZ domain-containing proteins is independent of p53 degradation or hTERT activation but does correlate with oncogenic potential (R. A. Watson, M. Thomas, L. Banks, and S. Roberts, J. Cell Sci. 116:4925-4934, 2003) and is essential for induction of epithelial hyperplasia in vivo (M. L. Nguyen, M. M. Nguyen, D. Lee, A. E. Griep, and P. F. Lambert, J. Virol. 77:6957-6964, 2003). In this study, we found that HPV type 16 E6 was able to activate NF-κB in airway epithelial cells through the induction of nuclear binding activity of p52-containing NF-κB complexes in a PDZ binding motif-dependent manner. Transcript accumulation for the NF-κB-responsive antiapoptotic gene encoding cIAP-2 and binding of nuclear factors to the proximal NF-κB binding site of the cIAP-2 gene promoter are induced by E6 expression. Furthermore, E6 is able to protect cells from TNF-induced apoptosis. All of these E6-dependent phenotypes are dependent on the presence of the PDZ binding motif of E6. Our results imply a role for targeting of PDZ proteins by E6 in NF-κB activation and protection from apoptosis in airway epithelial cells.
Characterization of the Early Steps of Hepatitis C Virus Infection by Using Luciferase Reporter Viruses
Authors: Koutsoudakis George , Kaul Artur , Steinmann Eike , Kallis Stephanie , Lohmann Volker , Pietschmann Thomas , Bartenschlager Ralf
Published: May 2006

The lack of an efficient system to produce hepatitis C virus (HCV) particles has impeded the analysis of the HCV life cycle. Recently, we along with others demonstrated that transfection of Huh7 hepatoma cells with a novel HCV isolate (JFH1) yields infectious viruses. To facilitate studies of HCV replication, we generated JFH1-based bicistronic luciferase reporter virus genomes. We found that RNA replication of the reporter construct was only slightly attenuated and that virus titers produced were only three- to fivefold lower compared to the parental virus, making these reporter viruses an ideal tool for quantitative analyses of HCV infections. To expand the scope of the system, we created two chimeric JFH1 luciferase reporter viruses with structural proteins from the Con1 (genotype 1b) and J6CF (genotype 2a) strains. Using these and the authentic JFH1 reporter viruses, we analyzed the early steps of the HCV life cycle. Our data show that the mode of virus entry is conserved between these isolates and involves CD81 as a key receptor for pH-dependent virus entry. Competition studies and time course experiments suggest that interactions of HCV with cell surface-resident glycosaminoglycans aid in efficient infection of Huh7 cells and that CD81 acts during a postattachment step. The reporter viruses described here should be instrumental for investigating the viral life cycle and for the development of HCV inhibitors.
Kaposi's Sarcoma-Associated Herpesvirus Lytic Gene ORF57 Is Essential for Infectious Virion Production
Authors: Han Zhao , Swaminathan Sankar
Published: May 2006

The ORF57 gene of Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a nuclear protein expressed during the lytic phase of KSHV replication. An ORF57 homolog is present in all known human herpesviruses and many animal herpesviruses. Many of these proteins have been demonstrated to have essential transcriptional and posttranscriptional regulatory functions. ORF57 enhances expression of reporter genes posttranscriptionally in vitro and may synergize with transcription factors to enhance gene transcription. However, the biologic role of ORF57 in KSHV replication has not been established. In this study, we demonstrate that ORF57 is essential for productive KSHV lytic replication by constructing a recombinant KSHV in which ORF57 expression has been specifically inactivated. The ORF57-null KSHV recombinant was unable to produce virion progeny or fully express several other lytic KSHV genes except when ORF57 was provided in
trans
. The Epstein-Barr virus (EBV) homolog of ORF57, SM, was unable to rescue lytic KSHV virion production, although EBV SM does enhance KSHV lytic gene expression from the ORF57-null mutant. Conversely, ORF57 did not rescue an SM-null recombinant EBV, indicating the existence of virus-specific functions for the ORF57 family of genes.
Extraintestinal Spread and Replication of a Homologous EC Rotavirus Strain and a Heterologous Rhesus Rotavirus in BALB/c Mice
Authors: Fenaux M. , Cuadras M. A. , Feng N. , Jaimes M. , Greenberg H. B.
Published: May 2006

Although rotavirus infection has generally been felt to be restricted to the gastrointestinal tract, over the last two decades there have been sporadic reports of children with acute or fatal cases of rotavirus gastroenteritis testing positive for rotavirus antigen and/or nucleic acid in various extraintestinal locations such as serum, liver, kidney, bladder, testes, nasal secretions, cerebrospinal fluid, and the central nervous system. Recently, studies in animals and people have demonstrated that rotavirus antigenemia is a common event during natural infection. In this study, we extend these observations and compare the intestinal and extraintestinal spread of wild-type homologous murine rotavirus EC and a heterologous strain, rhesus rotavirus (RRV), in newborn mice. A strand-specific quantitative reverse transcription-PCR (ssQRT-PCR) assay was used to quantify the ability of different rotavirus strains to spread and replicate extraintestinally. Both strain EC and RRV were detected extraintestinally in the mesenteric lymph nodes (MLN), livers, lungs, blood, and kidneys. Extraintestinal replication, as measured by ssQRT-PCR, was most prominent in the MLN and occurred to a lesser degree in the livers, kidneys, and lungs. In the MLN, strain EC and RRV had similar (
P
< 0.05) RNA copy numbers, although EC was present at a 10,000-fold excess over RRV in the small intestine. Rotavirus nonstructural protein 4 (NSP4) and/or assembled triple-layered particles, indicated by immunostaining with the VP7 conformation-dependent monoclonal antibody 159, were detected in the MLN, lungs, and livers of EC- and RRV-inoculated mice, confirming the ssQRT-PCR findings. Infectious RRV was detected in the MLN in quantities exceeding the amount present in the small intestines or blood. The cells in the MLN that supported rotavirus replication included dendritic cells and potentially B cells and macrophages. These data indicate that extraintestinal spread and replication occurs commonly during homologous and some heterologous rotaviral infections; that the substantial host range restrictions for rhesus rotavirus, a heterologous strain present in the intestine, are not necessarily apparent at systemic sites; that the level and location of extraintestinal replication varies between strains; that replication can occur in several leukocytes subsets; and that extraintestinal replication is likely a part of the normal pathogenic sequence of homologous rotavirus infection.
Epitopes Expressed in Different Adenovirus Capsid Proteins Induce Different Levels of Epitope-Specific Immunity
Authors: Krause Anja , Joh Ju H. , Hackett Neil R. , Roelvink Peter W. , Bruder Joseph T. , Wickham Thomas J. , Kovesdi Imre , Crystal Ronald G. , Worgall Stefan
Published: May 2006

On the basis of the concept that the capsid proteins of adenovirus (Ad) gene transfer vectors can be genetically manipulated to enhance the immunogenicity of Ad-based vaccines, the present study compared the antiantigen immunogenicity of Ad vectors with a common epitope of the hemagglutinin (HA) protein of the influenza A virus incorporated into the outer Ad capsid protein hexon, penton base, fiber knob, or protein IX. Incorporation of the same epitope into the different capsid proteins provided insights into the correlation between epitope position and antiepitope immunity. Following immunization of three different strains of mice (C57BL/6, BALB/c, and CBA) with either an equal number of Ad particles (resulting in a different total HA copy number) or different Ad particle numbers (to achieve the same HA copy number), the highest primary (immunoglobulin M [IgM]) and secondary (IgG) anti-HA humoral and cellular CD4 gamma interferon and interleukin-4 responses against HA were always achieved with the Ad vector carrying the HA epitope in fiber knob. These observations suggest that the immune response against an epitope inserted into Ad capsid proteins is not necessarily dependent on the capsid protein number and imply that the choice of incorporation site in Ad capsid proteins in their use as vaccines needs to be compared in vivo.
Regulation of the EBNA1 Epstein-Barr Virus Protein by Serine Phosphorylation and Arginine Methylation
Authors: Shire Kathy , Kapoor Priya , Jiang Ke , Hing Margaret Ng Thow , Sivachandran Nirojini , Nguyen Tin , Frappier Lori
Published: May 2006

The Epstein-Barr virus (EBV) EBNA1 protein is important for the replication and mitotic segregation of EBV genomes in latently infected cells and also activates the transcription of some of the viral latency genes. A Gly-Arg-rich region between amino acids 325 and 376 is required for both the segregation and transcriptional activation functions of EBNA1. Here we show that this region is modified by both arginine methylation and serine phosphorylation. Mutagenesis of the four potentially phosphorylated serines in this region indicated that phosphorylation of multiple serines contributes to the efficient segregation of EBV-based plasmids by EBNA1, at least in part by increasing EBNA1 binding to hEBP2. EBNA1 was also found to bind the arginine methyltransferases PRMT1 and PRMT5. Multiple arginines in the 325-376 region were methylated in vitro by PRMT1 and PRMT5, as was an N-terminal Gly-Arg-rich region between amino acids 41 and 50. EBNA1 was also shown to be methylated in vivo, predominantly in the 325-376 region. Treatment of cells with a methylation inhibitor or down-regulation of PRMT1 altered EBNA1 localization, resulting in the formation of EBNA1 rings around the nucleoli. The results indicate that EBNA1 function is influenced by both serine phosphorylation and arginine methylation.
Characterization of chromosome-specific S-SAP markers and their use in studying genetic diversity in Aegilops species
Authors: Nagy Ervin D , Molnár István , Schneider Annamária , Kovács Géza , Molnár-Láng Márta
Published: May 2006

The short interspersed nuclear element (SINE), Au, was used to develop sequence-specific amplified polymorphism (S-SAP) markers for U- and M-genome chromosomes. The markers were localized using Triticum aestivum (wheat) – Aegilops geniculata and wheat – Aegilops biuncialis disomic chromosome addition lines. Thirty-seven markers distributed over 6 U and 6 M chromosomes were produced. A genetic diversity study carried out on 37 accessions from Ae. biuncialis, Ae. comosa, Ae. geniculata, and Ae. umbellulata suggested that Ae. biuncialis have arisen from its diploid ancestors more recently than Ae. geniculata. Several earlier studies indicated that the M genomes in polyploid Aegilops species had accumulated substantial rearrangements, whereas the U genomes remained essentially unmodified. However, this cannot be attributed to the preferential insertion of retroelements into the M genome chromosomes. Fourteen markers from a total of 8 chromosomes were sequenced; 3 markers were similar to known plant genes, 1 was derived from a long terminal repeat (LTR) retrotransposon, and 10 markers did not match to any known DNA sequences, suggesting that they were located in the highly variable intergenic regions.Key words: Aegilops, U and M genomes, S-SAP, genetic diversity, sequence characterization.
Quantification of the retrotransposon BARE-1 reveals the dynamic nature of the barley genome
Authors: Soleimani V D , Baum B R , Johnson D A
Published: May 2006

We used quantitative real-time PCR analysis to measure the copy number of the BARE-1 retrotransposon in 5 cultivars of barley (Hordeum vulgare), as well as in samples from its wild relative, Hordeum spontaneum. Two sets of PCR primers were used to amplify regions within the long terminal repeat (LTR) and the reverse transcriptase (RT) gene of BARE-1 (GenBank accession Z17327). The LTR primers detected an average of 2.148 × 105 ± 0.012 × 105 copies per haploid genome among barley samples, whereas the RT primers detected an average of 1.588 × 104 ± 0.085 × 104 copies. The average ratio of LTR:RT was estimated to be 13.5:1. This finding indicates that more than 7% of the barley genome is occupied by BARE-1 elements in the form of solo LTRs and another 2.6% of the genome is occupied by the full-length element. Taken together, BARE-1 sequences represent approximately 9.6% of the barley genome among the barley plants used in this study. For the above estimation, a genome size of 5.44 × 103 Mb for H. vulgare and 5.39 × 103 Mb for H. spontaneum were assumed. Our study on quantification results of the BARE-1 for a small group of barley cultivars showed that there are significant differences among cultivars in terms of BARE-1 copy number, providing further evidence that BARE-1 is active and has a major role in shaping the barley genome as a result of breeding and selection. Quantification results also showed that most of the elements (> 90%) are present as truncated copies (solo LTRs). These results show that there is a high level of recombination leading to the formation of truncated elements and a subsequent DNA loss from the genome. Taken together, our study provides a glimpse into a dynamic micro-evolutionary process that is the by-product of genome reshuffling and directional selection in barley breeding programsKey words: BARE-1, genome evolution, quantification, real-time PCR, retrotransposons.
A framework linkage map of perennial ryegrass based on SSR markers
Authors: Gill G P , Wilcox P L , Whittaker D J , Winz R A , Bickerstaff P , Echt C E , Kent J , Humphreys M O , Elborough K M , Gardner R C
Published: May 2006

A moderate-density linkage map for Lolium perenne L. has been constructed based on 376 simple sequence repeat (SSR) markers. Approximately one third (124) of the SSR markers were developed from GeneThresher®libraries that preferentially select genomic DNA clones from the gene-rich unmethylated portion of the genome. The remaining SSR marker loci were generated from either SSR-enriched genomic libraries (247) or ESTs (5). Forty-five percent of the GeneThresher SSRs were associated with an expressed gene. Unlike EST-derived SSR markers, GeneThresher SSRs were often associated with genes expressed at a low level, such as transcription factors. The map constructed here fulfills 2 definitions of a "framework map". Firstly, it is composed of codominant markers to ensure map transferability either within or among species. Secondly, it was constructed to achieve a level of statistical confidence in the support-for-order of marker loci. The map consists of 81 framework SSR markers spread over 7 linkage groups, the same as the haploid chromosome number. Most of the remaining 295 SSR markers have been placed into their most likely interval on the framework map. Nine RFLP markers and 1 SSR marker from another map constructed using the same pedigree were also incorporated to extend genome coverage at the terminal ends of 5 linkage groups. The final map provides a robust framework with which to conduct investigations into the genetic architecture of trait variation in this commercially important grass species.Key words: Framework map, perennial ryegrass, SSR, simple sequence repeat, GeneThresher, Lolium perenne.
Electron transport pathways for the oxidation of endogenous substrate(s) inAcidithiobacillus ferrooxidans
Authors: Chen Yongqiang , Suzuki Isamu
Published: May 2006

Oxidation of endogenous substrate(s) of Acidithiobacillus ferrooxidans with O2or Fe3+as electron acceptor was studied in the presence of uncouplers and electron transport inhibitors. Endogenous substrate was oxidized with a respiratory quotient (CO2produced/O2consumed) of 1.0, indicating its carbohydrate nature. The oxidation was inhibited by complex I inhibitors (rotenone, amytal, and piericidin A) only partially, but piericidin A inhibited the oxidation with Fe3+nearly completely. The oxidation was stimulated by uncouplers, and the stimulated activity was more sensitive to inhibition by complex I inhibitors. HQNO (2-heptyl-4-hydroxyquinoline N-oxide) also stimulated the oxidation, and the stimulated respiration was more sensitive to KCN inhibition than uncoupler stimulated respiration. Fructose, among 20 sugars and sugar alcohols including glucose and mannose, was oxidized with a CO2/O2ratio of 1.0 by the organism. Iron chelators in general stimulated endogenous respiration, but some of them reduced Fe3+chemically, introducing complications. The results are discussed in view of a branched electron transport system of the organism and its possible control.Key words: Acidithiobacillus ferrooxidans, endogenous respiration, uncouplers, electron transport.
A comparison of selected methods for measuring the virulence properties ofListeriaspp.
Authors: Chiu Sally , Vanderlinde Paul B , Dykes Gary A
Published: May 2006

The comparative ability of different methods to assess virulence of Listeria species was investigated in ten Listeria strains. All strains were initially subjected to pulsed-field gel electrophoresis analysis to determine their relatedness. Virulence characteristics were subsequently tested for by (i) determining the presence of six virulence genes by polymerase chain reaction; (ii) testing for the production of listeriolysin O, phosphatidylcholine phospholipase C, and phosphatidylinositol-specific phospholipase C; (iii) investigating the hydrophobicity of the strains; (iv) determining the strains ability to attach to, enter, and replicate within the Caco-2 cells. Variations in most of the virulence characteristics were obvious across the strains for the range of tests performed. A wide range of anomalous results among methods were apparent. In particular, the presence of virulence genes was found to be unrelated to the production of virulence-associated proteins in vitro, while virulence protein production and hydrophobicity in Listeria monocytogenes were found to be unrelated or marginally related, respectively, to the ability to invade the Caco-2 cell line. It was concluded that the methods investigated were unable to consistently and unequivocally measure the differences in the virulence properties of the strains.Key words: Listeria monocytogenes, virulence, pathogenicity.
Foot odor due to microbial metabolism and its control
Authors: Ara Katsutoshi , Hama Masakatsu , Akiba Syunichi , Koike Kenzo , Okisaka Koichi , Hagura Toyoki , Kamiya Tetsuro , Tomita Fusao
Published: May 2006

To characterize foot odor, we analyzed its components by sensory tests, isolated microorganisms that produce it, and evaluated the mechanism of the occurrence of foot odor. As a result, foot odor was found to be derived from isovaleric acid, which is produced when Staphylococcus epidermidis, a resident species of the normal cutaneous microbial flora, degrades leucine present in sweat. In addition, Bacillus subtilis was detected in the plantar skin of subjects with strong foot odor, and this species was shown to be closely associated with increased foot odor. Therefore, we screened various naturally occurring substances and fragrant agents that inhibit microbial production of foot odor without disturbing the normal microbial flora of the human skin. As a result, we identified citral, citronellal, and geraniol as fragrant agents that inhibit the generation of isovaleric acid at low concentrations.Key words: foot odor, microbial metabolism, isovaleric acid, leucine dehydrogenase.
Antimicrobial compounds produced byActinomadurasp. AC104 isolated from an Algerian Saharan soil
Authors: Badji B , Zitouni A , Mathieu F , Lebrihi A , Sabaou N
Published: May 2006

During a search for nonpolyenic antifungal antibiotics, an actinomycete designated AC104 was isolated from a Saharan soil sample by a dilution agar plating method using a chitin - vitamins B medium supplemented with rifampicin. Isolate AC104 presented the morphological and the chemical characteristics of the genus Actinomadura. On the basis of 76 physiological tests and 16S rDNA analysis, this isolate was determined to be quite different from the known species of Actinomadura. It is active against filamentous fungi and both Gram-positive and Gram-negative bacteria. The production of antibiotic substances was investigated using several culture media. The highest antimicrobial activities were obtained on ISP2 medium. The benzenic extract contained five bioactive spots detected on thin layer chromatography plates. Among these antibiotics, a complex called 104A, which showed the more interesting antifungal activity, was selected and purified by reverse-phase high-pressure liquid chromatography. This complex is composed of four compounds. Ultraviolet-visible, infrared, mass, and1H nuclear magnetic resonance spectroscopy studies showed that these molecules contain an aromatic ring substituted by aliphatic chains. These compounds differ from the known antibiotics produced by Actinomadura species.Key words: Actinomadura, taxonomy, antimicrobial compounds, Saharan soil.
Extracytoplasmic storage as the nickel resistance mechanism in a natural isolate ofPseudomonas putidaS4
Authors: Tripathi V N , Srivastava S
Published: May 2006

Metal resistances in microbes are important to study not only to understand metal homeostasis but also to use such organisms further in environmental bioremediation. Nickel (Ni2+) is an important micronutrient, which at higher concentration becomes toxic. Many Ni2+-resistant organisms are known, which resist metal by active efflux. Pseudomonas putida S4, a natural isolate from India, is reported to show a multi-metal resistance profile. In the present study, the Ni2+-resistance mechanism in strain S4 was examined. Wild-type cells gradually accumulated Ni2+but kept it preferentially in the periplasmic space in a bound form. In Ni2+-sensitive mutants, periplasmic storage was disturbed and more metal accumulated cytoplasmically, producing toxicity. Sodium dodecyl sulphate – polyacrylamide gel electrophoresis analysis of periplasmic proteins revealed a band of approximately 18 kDa, which appeared only in Ni2+-exposed wild-type cells, and which was absent from cells not exposed to Ni2+as well as from Ni2+-sensitive mutants. On the basis of these observations, we propose a Ni2+-resistance mechanism in P. putida S4 based on sequestration of metal in the periplasmic space. This is the first study of sequestration-based Ni2+resistance.Key words: nickel, resistance, periplasmic storage, Pseudomonas putida.
Development of AFLP-derived, functionally specific markers for environmental persistence studies of fungal strains
Authors: Hynes S S , Chaudhry O , Providenti M A , Smith M L
Published: May 2006

The ability to rapidly identify and quantify a microbial strain in a complex environmental sample has widespread applications in ecology, epidemiology, and industry. In this study, we describe a rapid method to obtain functionally specific genetic markers that can be used in conjunction with standard or real-time polymerase chain reaction (PCR) to determine the abundance of target fungal strains in selected environmental samples. The method involves sequencing of randomly cloned AFLP (amplified fragment length polymorphism) products from the target organism and the design of PCR primers internal to the AFLP fragments. The strain-specific markers were used to determine the fate of three industrially relevant fungi, Aspergillus niger, Aspergillus oryzae, and Chaetomium globosum, during a 4 month soil microcosm experiment. The persistence of each of the three fungal strains inoculated separately into intact soil microcosms was determined by PCR analyses of DNA directly extracted from soil. Presence and absence data based on standard PCR and quantification of the target DNA by real-time PCR showed that all three strains declined after inoculation (~14-, 32-, and 4-fold for A. niger, A. oryzae, and C. globosum, respectively) but remained detectable at the end of the experiment, suggesting that these strains would survive for extended periods if released into nature.Key words: Canada domestic substances list (DSL), Canadian Environmental Protection Act (CEPA), genetically modified organisms (GMO), quantitative polymerase chain reaction (qPCR).
Model plants for studying the interaction between Methylobacterium mesophilicum and Xylella fastidiosa
Authors: Andreote Fernando D , Lacava Paulo T , Gai Cláudia S , Araújo Welington L , Maccheroni, Jr. Walter , van Overbeek Leonard S , van Elsas Jan Dirk , Azevedo João Lúcio
Published: May 2006

Over the last few years, endophytic bacterial communities associated with citrus have been studied as key components interacting with Xylella fastidiosa. In this study, we investigated the possible interaction between the citrus endophyte Methylobacterium mesophilicum SR1.6/6 and X. fastidiosa in model plants such as Catharanthus roseus (Madagaskar periwinkle) and Nicotiana clevelandii (Clevelands tobacco). The aim of this study was to establish the fate of M. mesophilicum SR1.6/6 after inoculation of C. roseus and N. clevelandii plants, using PCR–DGGE (polymerase chain reaction – denaturing gradient gel electrophoresis) and plating techniques. Shifts in the indigenous endophytic bacterial communities were observed in plants inoculated with strain SR1.6/6, using specific primers targeting α- and β-Proteobacteria. Cells of strain SR1.6/6 were observed in a biofilm structure on the root and hypocotyl surfaces of in vitro seedlings inoculated with M. mesophilicum SR1.6/6. This emphasizes the importance of these tissues as main points of entrance for this organism. The results showed that C. roseus and N. clevelandii could be used as model plants to study the interaction between M. mesophilicum and X. fastidiosa.Key words: endophytic, Methylobacterium, model plants, DGGE.
Variable adhesion and diurnal population patterns of epiphytic yeasts on creeping bentgrass
Authors: Allen Tom W , Burpee Leon L , Buck James W
Published: May 2006

Irrigation and an in vitro agitation assay were used to determine the percentage of the epiphytic yeast community (Cryptococcus, Pseudozyma, Rhodotorula, and Sporobolomyces) adhering to the phylloplane of creeping bentgrass (Agrostis palustris (Huds.) Pers.). Colony-forming units (cfu) of total epiphytic yeast populations (adherent and nonadherent cells) and of adherent populations (cells not removed by agitation) were determined by leaf washing and dilution plating. In an in vitro assay, 40.0% and 57.1% of the yeast adhered to the leaves, whereas, in initial field trials the percentage of adherent yeasts ranged from 40.0% to 71.9% of the total population. Adherent yeast cfu on leaves in the morning were significantly lower on bentgrass (8.0 × 103to 3.1 × 104cfu·cm–2) compared with total yeast cfu (1.4 × 104to 4.7 × 104cfu·cm–2) on the nonirrigated control. No differences in yeast populations were observed between irrigated and nonirrigated plots 2 h after the 0900 treatments. Yeast populations followed a diurnal pattern, with larger cfu recovered from bentgrass leaves in the morning and significantly lower populations recovered in the afternoon. At 1400 the adherent yeast were 83.1%–100% of the total yeast population recovered from the leaves. The relative adhesiveness of the epiphytic yeast community on bentgrass leaves is dynamic with nonadherent cells making up a larger percentage of the population in the mornings than the afternoons.Key words: adherence, Cryptococcus, leaf surface, Rhodotorula, turfgrass.
Identification of low-temperature-regulated genes in the fire blight pathogenErwinia amylovora
Authors: Goyer Claudia , Ullrich Matthias S
Published: May 2006

Genes involved in pathogenicity of several plant pathogens were shown to be induced at relatively cold temperatures. Loci from the fire blight pathogen Erwinia amylovora (Burrill) induced at 18 °C were identified using the miniTn5 transposon that contains the promoterless reporter gene gusA coding for β-glucuronidase (GUS). Certain mutants (2.7%) expressed GUS predominantly at 18 °C on minimal medium plates, indicating that the transposon had been inserted downstream of a putatively thermoregulated promoter. Those mutants were further screened with a quantitative GUS fluorometric assay. A total of 21 mutants were selected: 19 mutants had a transposon insertion in temperature-dependent genetic loci, with a 2.2- to 6.3-fold induction of gusA gene expression at 18 °C, and two mutants with impaired growth at 18 °C. Some of these genetic loci encoded (i) proteins implicated in flagella biosynthesis, biotin biosynthesis, multi-drug efflux, and type II secretion protein, and (ii) proteins of unknown function.Key words: fire blight, Erwinia amylovora, transposon mutagenesis, gene regulation, low temperature.
Culture conditions for the production of an acidic exopolysaccharide by the nitrogen-fixing bacterium Burkholderia tropica
Authors: Serrato Rodrigo V , Sassaki Guilherme L , Cruz Leonardo M , Pedrosa Fábio O , Gorin Philip A.J , Iacomini Marcello
Published: May 2006

The endophytic diazotrophic bacterium Burkholderia tropica, strain Ppe8, produced copious amounts of exopolysaccharide (EPS) on batch growth in liquid synthetic media containing mannitol and glutamate as carbon and nitrogen sources. The effect of various aeration regimes and carbon source concentrations on EPS production was determined, as well as the effects of temperature and time of growth. The degree of aeration had a great influence on the yield of EPS, in contrast with the C:N ratio of the medium. Growth temperature also affected the EPS yield after the first 24 h of culture but seemed to be irrelevant after that. After isolation and purification, the EPS was analyzed by high-performance size exclusion chromatography and multiangle laser light scattering (HPSEC–MALLS), revealing a molecular mass of 300 kDa. The acid hydrolyzate of EPS was examined by HPLC and found to contain Glc, Rha, GlcA, and an aldobiouronic acid. The latter was found to have a GlcA and a Rha unit. Carboxy-reduced EPS contained Glc and Rha (3:2). The monosaccharide composition of the native acidic EPS was calculated as GlcA, Glc, and Rha in a molar ratio of 1:2:2.Key words: Burkholderia, endophyte, diazotrophic, exopolysaccharide, EPS.
Modification of cutaneous vasodilator response to heat stress by daytime exogenous melatonin administration
Authors: Aoki Ken , Stephens Dan P. , Zhao Kun , Kosiba Wojciech A. , Johnson John M.
Published: May 2006

In humans, the nocturnal fall in internal temperature is associated with increased endogenous melatonin and with a shift in the thermoregulatory control of skin blood flow (SkBF), suggesting a role for melatonin in the control of SkBF. The purpose of this study was to test whether daytime exogenous melatonin would shift control of SkBF to lower internal temperatures during heat stress, as is seen at night. Healthy male subjects ( n = 8) underwent body heating with melatonin administration (Mel) or without (control), in random order at least 1 wk apart. SkBF was monitored at sites pretreated with bretylium to block vasoconstrictor nerve function and at untreated sites. Cutaneous vascular conductance, calculated from SkBF and arterial pressure, sweating rate (SR), and heart rate (HR) were monitored. Skin temperature was elevated to 38°C for 35–50 min. Baseline esophageal temperature (Tes) was lower in Mel than in control ( P < 0.01). The Tes threshold for cutaneous vasodilation and the slope of cutaneous vascular conductance with respect to Tes were also lower in Mel at both untreated and bretylium-treated sites ( P < 0.05). The Tes threshold for the onset of sweating and the Tes for a standard HR were reduced in Mel. The slope of the relationship of HR, but not SR, to Tes was lower in Mel ( P < 0.05). These findings suggest that melatonin affects the thermoregulatory control of SkBF during hyperthermia via the cutaneous active vasodilator system. Because control of SR and HR are also modified, a central action of melatonin is suggested.
Increased Brain Expression of Matrix Metalloproteinase-9 After Ischemic and Hemorrhagic Human Stroke
Authors: Rosell Anna , Ortega-Aznar Arantxa , Alvarez-Sabín José , Fernández-Cadenas Israel , Ribó Marc , Molina Carlos A. , Lo Eng H. , Montaner Joan
Published: May 2006

Background and Purpose—

Abnormal expression of some matrix metalloproteinases (MMP) has shown to play a deleterious role in brain injury in experimental models of cerebral ischemia. We aimed to investigate MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in brain parenchyma in both ischemic and hemorrhagic strokes.

Methods—

Postmortem fresh brain tissue from 6 ischemic and 8 hemorrhagic stroke patients was obtained within the first 6 hours after death. Finally, 78 brain tissue samples from different areas (infarct, peri-infarct, perihematoma and contralateral hemisphere) were studied. To quantify gelatinase content we performed gelatin zymograms that were confirmed by Western Blot Analysis, immunohistochemistry to localize MMP source, and in situ zymography to detect gelatinase activity.

Results—

Among ischemic cases, gelatin zymography showed increased MMP-9 content in infarct core although peri-infarct tissue presented also higher levels than contralateral hemisphere (
P
<0.0001 and
P
=0.042, respectively). Within infarct core, MMP-9 was mainly located around blood vessels, associated to neutrophil infiltration and activated microglial cells. In peri-infarct areas the major source of MMP-9 were microglial cells. Tissue around intracranial hemorrhage also displayed higher MMP-9 levels than contralateral hemisphere (
P
=0.008) in close relationship with glial cells. MMP-2 was constitutively expressed and remained invariable in different brain areas.

Conclusions—

Our results demonstrate in situ higher levels of MMP-9 in human brain tissue after ischemic and hemorrhagic stroke, suggesting a contribution of MMP-9 to ischemic brain injury and perihematoma edema.
Liquor Is Quicker
Authors: Lanza-Kaduce Lonn , Capece Michael , Alden Helena
Published: May 2006

Akers’s Social Structure-Social Learning Theory (SS-SL) has recently been criticized for dealing with social structural factors as exogenous variables rather than integrating them into propositions that specify the learning processes better. For example, Morash has argued that SS-SL has largely ignored the ways in which gender structures institutions, interactions, and behaviors and takes Akers to task for ignoring feminist theories. This research uses Core Alcohol and Drug Survey data from a subsample of White unmarried college students from eight diverse campuses throughout the United States to examine how conflict structured by gender affects differences in the use of alcohol before sex. The research specifically examines Akers’s claim that social learning variables (in this case, anticipated reward and risk of harm) will substantially mediate the effects of structural variables rather than modulate or moderate them. The policy implications of the research are discussed.
Developing a physicians’ palliative care pain hotline in Maryland
Authors: Gloth F. Michael , Schwartz Jack
Published: May 2006

Physicians have had relatively little formal training in pain management and palliative care. For this reason, a telephone consultation service was offered, the physicians’ palliative care pain hotline, that would allow physicians to call a toll-free number and, within 15 minutes, speak to a board-certified physician in hospice and palliative medicine. To our knowledge, this is the first program of its kind. This article describes the process involved in creating such a pain hotline and reports on some data collected on its use in the first 10 months. This report should help others who have an interest in establishing a similar program.
Self-esteem in a palliative care population
Authors: Babar Mahrukh , ul Haq Sheikh Moeen
Published: May 2006

Objective. To study the effects of palliative care treatment in cancer patients, on their self-esteem.

Design and setting. It was a cross-sectional study conducted at Shaukat Khanum Memorial Cancer Hospital and Research Center, Lahore, Pakistan (SKMCH & RC).

Participants. A total of 60 referred cancer patients receiving palliative care were interviewed using the Mehrabian self-esteem scale (MSE). No follow-up was required. All the patients were over 16 years of age. The sample was not restricted to any one type of cancer. Since lack of education could be a contributing factor, only patients who had a minimum of an eighth-grade education were included in the study.

Results. Four variables were studied: gender, age, diagnostic-related group, and performance status. Performance status was determined using the Eastern Cooperative Oncology Group Scale (ECOG). The only significant correlation was between self-esteem and performance status, with a p value of 0.04.

Conclusion. We conclude that the patients who are functionally active (not disabled) have a higher level of self-esteem, as compared to those who are disabled. This is quite understandable, as the patients who are not dependent on others for their needs feel worthy, resulting in higher self-esteem. This was the first study of its kind done in Pakistan. We identified two limitations. First, the sample size was small. Second, although the Mehrabian self-esteem scale is a valid, reliable, and standardized scale, it was not standardized for the population of Pakistan.