bluuebunny/crossref_metadata_2025 huggingface dataset:
Using the Psychology of Evil To Do Good
Authors: Miller Greg
Published: April 2011

Forty years after the infamous Stanford Prison Experiment, Phil Zimbardo thinks he can apply its lessons to teach ordinary people to be heroes.
Jeremy Berg: An Independent Scientist Departs NIH's Ranks
Authors: Kaiser Jocelyn
Published: April 2011

The outgoing director of the basic research institute at the U.S. National Institutes of Health has won scientists' hearts and minds by championing transparency.
Ancient Footprints Tell Tales of Travel
Authors: Gibbons Ann
Published: April 2011

By scrutinizing a 120,000-year-old trail of footprints, researchers have gotten their first snapshot of what a traveling group of archaic humans looked like, according to paleoanthropologists reporting at the American Association of Physical Anthropologists meeting.
Snapshots From the Meeting
Authors: Gibbons Ann
Published: April 2011

Snapshots from the American Association of Physical Anthropologists meeting include a piece of a thighbone that fits perfectly on top of a broken bone found 29 years earlier and three new wrist bones from a second individual of the diminutive "Hobbit" from the Indonesian island of Flores.
Massilia timonae Infection Presenting as Generalized Lymphadenopathy in a Man Returning to Belgium from Nigeria
Authors: Van Craenenbroeck Amaryllis H. , Camps Kim , Zachée Pierre , Wu Ka Lung
Published: April 2011

We report a case of apparent malaria infection presented with a syndrome of painless, generalized lymphadenopathy without granulomas shortly after exposure to fresh water in rural West Africa. Residual infection with
Massilia timonae
was diagnosed and successfully treated with co-trimoxazole.
Acinetobacter soli as a Cause of Bloodstream Infection in a Neonatal Intensive Care Unit
Authors: Pellegrino Flávia Lúcia Piffano Costa , Vieira Verônica V. , Baio Paulo Victor Pereira , dos Santos Rosana Maria R. , dos Santos Ana Lucia Alves , Santos Nadir Gomes de Barros , Meohas Martha Maria Gaudie Ley , Santos Rodrigo Teixeira , de Souza Talita Coelho , da Silva Dias Rubens Clayton , Santoro-Lopes Guilherme , Riley Lee W. , Moreira Beatriz Meurer
Published: April 2011

Acinetobacter soli
is a new bacterial species described from forest soil. Five cases of bloodstream infection caused by
A. soli
clonal isolates are reported here for the first time. The patients were neonates admitted to an intensive care unit. This is a new neonatal pathogen with the potential to cause outbreaks.
Cutaneous Diphtheria in the Urban Poor Population of Vancouver, British Columbia, Canada: a 10-Year Review
Authors: Lowe C. F. , Bernard K. A. , Romney M. G.
Published: April 2011

Between 1998 and 2007, records from 33 patients with cutaneous diphtheria from Vancouver's inner city were reviewed. Cases were associated with injection drug use and poverty. Coinfections with
Staphylococcus aureus
,
Streptococcus pyogenes
, and
Arcanobacterium haemolyticum
occurred.
Corynebacterium diphtheriae
is endemic in Vancouver's urban core, with strains of multilocus sequence type (MLST) 76 predominating.
Comparison of Automated Processing of Flocked Swabs with Manual Processing of Fiber Swabs for Detection of Nasal Carriage of
Staphylococcus aureus
Authors: Jones Gillian , Matthews Roger , Cunningham Richard , Jenks Peter
Published: April 2011

The sensitivity of automated culture of
Staphylococcus
aureus
from flocked swabs versus that of manual culture of fiber swabs was prospectively compared using nasal swabs from 867 patients. Automated culture from flocked swabs significantly increased the detection rate, by 13.1% for direct culture and 10.2% for enrichment culture.
Comparison of the Flotac-400 Dual Technique and the Formalin-Ether Concentration Technique for Diagnosis of Human Intestinal Protozoon Infection
Authors: Becker Sören L. , Lohourignon Laurent K. , Speich Benjamin , Rinaldi Laura , Knopp Stefanie , N'Goran Eliézer K. , Cringoli Giuseppe , Utzinger Jürg
Published: April 2011

There is a need for accurate diagnosis of intestinal parasite infections in humans, but currently available copromicroscopic techniques have shortcomings, such as low sensitivity. We compared the diagnostic accuracy of a further modified version of the recently developed Flotac technique with that of the widely used formalin-ether concentration technique (FECT) for the detection of intestinal protozoa in human stool samples. Formaldehyde-preserved stool samples from 108 individuals in Côte d'Ivoire were subjected to the Flotac-400 dual technique, using two different flotation solutions (FS), and to the FECT. Stool samples were examined according to computer-generated random lists by an experienced laboratory technician blinded for the results of each method. Both methods detected the same eight intestinal protozoon species. While the Flotac-400 dual technique (results from both FS combined) found higher prevalences of
Entamoeba coli
(77.8% versus 71.3%,
P
< 0.001),
Blastocystis hominis
(20.4% versus 13.0%,
P
= 0.458), and
Giardia intestinalis
(8.3% versus 6.5%,
P
< 0.001), the FECT detected higher prevalences of
Entamoeba histolytica/Entamoeba dispar
(27.8% versus 20.4%,
P
= 0.019) and four species of nonpathogenic intestinal protozoa. The diagnostic agreement between the two methods differed considerably depending on the intestinal protozoon investigated (Cohen's kappa measures; range, 0.01 to 0.46). Our study confirmed that the Flotac-400 dual technique can be utilized for the diagnosis of intestinal protozoon infections in humans. Since Flotac is a sensitive technique for the detection of soil-transmitted helminths and
Schistosoma mansoni
, it might become a viable copromicroscopic technique for the concurrent diagnosis of helminths and intestinal protozoon infections.
Clinical and Molecular Observations of Two Fatal Cases of Rotavirus-Associated Enteritis in Children in Italy
Authors: Medici Maria Cristina , Abelli Laura Anna , Martinelli Monica , Corradi Domenico , Dodi Icilio , Tummolo Fabio , Albonetti Valeria , Martella Vito , Dettori Giuseppe , Chezzi Carlo
Published: April 2011

Two fatal cases of infantile rotavirus enteritis occurred in northern Italy in 2005. Both children were severely dehydrated, and death was related to severe cerebral edema. Histological examination demonstrated extensive damage of the intestinal epithelium, villous atrophy or blunting, and macrophage infiltration. The two rotavirus strains were of the G1P[8] type and the long electropherotype. The 2005 G1P[8] rotaviruses differed in the NSP4, VP3, VP4, and VP7 genes from G1P[8] rotaviruses circulating in 2004, suggesting the onset of a new G1P[8] strain in the local population.
Shedding of Epstein-Barr Virus and Cytomegalovirus from the Genital Tract of Women in a Periurban Community in Andhra Pradesh, India
Authors: Silver Michelle I. , Paul Proma , Sowjanya Pavani , Ramakrishna Gayatri , Vedantham Haripriya , Kalpana Basany , Shah Keerti V. , Gravitt Patti E.
Published: April 2011

We found a large number of false-positive readings by visual inspection with acetic acid (VIA) in a study of cervical cancer screening strategies (VIA, human papillomavirus HPV DNA testing, and Pap cytology) in a periurban community in Andhra Pradesh, India. We evaluated whether these false-positive readings might be occurring as a result of infections with Epstein-Barr virus (EBV) or cytomegalovirus (CMV), prevalent latent herpesviruses known to be shed from the female genital tract. While we found that there was no association between VIA results and the presence of EBV or CMV in the cervix, we did find a high prevalence of both viruses: 20% for EBV and 26% for CMV. In multivariate analyses, CMV prevalence was associated with younger age, lack of running water in the home, and visually apparent cervical inflammation. EBV prevalence was associated with older age and a diagnosis of cervical intraepithelial neoplasia grade 1 or greater. The biological and clinical implications of these viruses at the cervix remain to be determined. The strong association between the presence of EBV and cervical disease warrants future exploration to determine whether EBV plays a causal role in disease development or if it is merely a bystander in the process.
Solobacterium moorei Bacteremia: Identification, Antimicrobial Susceptibility, and Clinical Characteristics
Authors: Pedersen Rune Micha , Holt Hanne Marie , Justesen Ulrik Stenz
Published: April 2011

We present five cases of
Solobacterium moorei
bacteremia. The isolates were identified with 16S rRNA gene sequencing and were susceptible to common antibiotics used for anaerobic infections. Bacteremia with
S. moorei
seems to be associated with debilitating conditions, but the prognosis of the infection appears to be good.
Sensitivity, Specificity, and Clinical Value of Human Papillomavirus (HPV) E6/E7 mRNA Assay as a Triage Test for Cervical Cytology and HPV DNA Test
Authors: Benevolo Maria , Vocaturo Amina , Caraceni Donatella , French Deborah , Rosini Sandra , Zappacosta Roberta , Terrenato Irene , Ciccocioppo Lucia , Frega Antonio , Rossi Paolo Giorgi
Published: April 2011

There is evidence that testing for human papillomavirus (HPV) E6/E7 mRNA is more specific than testing for HPV DNA. A retrospective study was carried out to evaluate the performance of the PreTect HPV-Proofer E6/E7 mRNA assay (Norchip) as a triage test for cytology and HPV DNA testing. This study analyzed 1,201 women, 688 of whom had a colposcopy follow-up and 195 of whom had histology-confirmed high-grade intraepithelial neoplasia or worse (CIN2+). The proportion of positive results and the sensitivity and specificity for CIN2+ were determined for HPV mRNA in comparison to HPV DNA and cytology. All data were adjusted for follow-up completeness. Stratified by cytological grades, the HPV mRNA sensitivity was 83% (95% confidence interval [CI] = 63 to 94%) in ASC-US (atypical squamous cells of undetermined significance), 62% (95% CI = 47 to 75%) in L-SIL (low-grade squamous intraepithelial lesion), and 67% (95% CI = 57 to 76%) in H-SIL (high-grade squamous intraepithelial lesion). The corresponding figures were 99, 91, and 96%, respectively, for HPV DNA. The specificities were 82, 76, and 45%, respectively, for HPV mRNA and 29, 13, and 4%, respectively, for HPV DNA. Used as a triage test for ASC-US and L-SIL, mRNA reduced colposcopies by 79% (95% CI = 74 to 83%) and 69% (95% CI = 65 to 74%), respectively, while HPV DNA reduced colposcopies by 38% (95% CI = 32 to 44%) and by 15% (95% CI = 12 to 19%), respectively. As a HPV DNA positivity triage test, mRNA reduced colposcopies by 63% (95% CI = 60 to 66%), having 68% sensitivity (95% CI = 61 to 75%), whereas cytology at the ASC-US+ threshold reduced colposcopies by 23% (95% CI = 20 to 26%), showing 92% sensitivity (95% CI = 87 to 95%). In conclusion, PreTect HPV-Proofer mRNA can serve as a better triage test than HPV DNA to reduce colposcopy referral in both ASC-US and L-SIL. It is also more efficient than cytology for the triage of HPV DNA-positive women. Nevertheless, its low sensitivity demands a strict follow-up of HPV DNA positive-mRNA negative cases.
The Human T-Cell Leukemia Virus Type 1 Oncoprotein Tax Controls Forkhead Box O4 Activity through Degradation by the Proteasome
Authors: Oteiza Alexandra , Mechti Nadir
Published: April 2011

Activation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway by the viral Tax oncoprotein plays a pivotal role in clonal expansion of human T-cell leukemia virus type 1 (HTLV-1)-infected cells. As the Forkhead box O (FoxO) tumor suppressors act as downstream effectors of PI3K/Akt, they represent good candidate targets whose dysregulation by Tax might be involved in HTLV-1-mediated activation and transformation of infected cells. In this report, we provide evidence showing that Tax induces a dose-dependent degradation of FoxO4 by the ubiquitin-proteasome pathway. Consistent with that, we demonstrate that Tax expression increases the interaction between FoxO4 and Mdm2 E3 ligase, leading to a strong FoxO4 polyubiquitination. These processes require the phosphorylation of FoxO4 by Akt, since a mutant of FoxO4 with mutations on its three Akt phosphorylation sites appears to be resistant to Tax-mediated degradation and ubiquitination. In addition, we show that Tax expression is associated with degradation and phosphorylation of endogenous FoxO4 in Jurkat T cells. Finally, we demonstrate that Tax represses FoxO4 transcriptional activity. Our study demonstrates that Tax can control FoxO4 protein stability and transcriptional activity and provides new insight into the subversion of cell signaling pathways during HTLV-1 infection.
Clathrin-Mediated Endocytosis in Living Host Cells Visualized through Quantum Dot Labeling of Infectious Hematopoietic Necrosis Virus
Authors: Liu Haibin , Liu Yi , Liu Shulin , Pang Dai-Wen , Xiao Gengfu
Published: April 2011

Infectious hematopoietic necrosis virus (IHNV) is an important fish pathogen that infects both wild and cultured salmonids. As a species of the genus
Novirhabdovirus
, IHNV is a valuable model system for exploring the host entry mechanisms of rhabdoviruses. In this study, quantum dots (QDs) were used as fluorescent labels for sensitive, long-term tracking of IHNV entry. Using live-cell fluorescence microscopy, we found that IHNV is internalized through clathrin-coated pits after the virus binds to host cell membranes. Pretreatment of host cells with chlorpromazine, a drug that blocks clathrin-mediated endocytosis, and clathrin light chain (LCa) depletion using RNA interference both resulted in a marked reduction in viral entry. We also visualized transport of the virus via the cytoskeleton (i.e., actin filaments and microtubules) in real time. Actin polymerization is involved in the transport of endocytic vesicles into the cytosol, whereas microtubules are required for the trafficking of clathrin-coated vesicles to early endosomes, late endosomes, and lysosomes. Disrupting the host cell cytoskeleton with cytochalasin D or nocodazole significantly impaired IHNV infectivity. Furthermore, infection was significantly affected by pretreating the host cells with bafilomycin A1, a compound that inhibits the acidification of endosomes and lysosomes. Strong colocalizations of IHNV with endosomes indicated that the virus is internalized into these membrane-bound compartments. This is the first report in which QD labeling is used to visualize the dynamic interactions between viruses and endocytic structures; the results presented demonstrate that IHNV enters host cells via clathrin-mediated endocytic, cytoskeleton-dependent, and low-pH-dependent pathways.
Delayed Cytosolic Exposure of Japanese Encephalitis Virus Double-Stranded RNA Impedes Interferon Activation and Enhances Viral Dissemination in Porcine Cells
Authors: Espada-Murao Lyre Anni , Morita Kouichi
Published: April 2011

Interferon is a principal component of the host antiviral defense system. In this study, abortive focus formation by Japanese encephalitis virus (JEV) in primate cells was accompanied by early interferon induction, while productive focus formation in porcine cells was associated with a late interferon response. Neutralization antibodies against interferon relieved the restricted infection in primate cells, and increasingly larger foci were generated as treatment with exogenous interferon was delayed, thereby establishing a solid correlation between interferon response and viral dissemination. However, delayed interferon induction in JEV-infected porcine cells occurred in the absence of active inhibition by the virus. We further demonstrated that JEV mediates interferon activation through double-stranded RNA and cytosolic pattern recognition receptors. Immunofluorescence and subcellular fractionation studies revealed that double-stranded RNA is concealed in intracellular membranes at an early phase of infection but eventually appears in the cytosol at later periods, which could then allow detection by cytosolic pattern recognition receptors. Interestingly, cytosolic exposure of double-stranded RNA was delayed in porcine cells compared to primate cells, independent of total double-stranded RNA levels and in correlation with the timing of the interferon response. Furthermore, when double-stranded RNA was artificially introduced into the cytosol of porcine cells, more rapid and robust interferon activation was triggered than in viral infection. Thus, cytosolic exposure of JEV double-stranded RNA is imperative for interferon induction, but in cell lines (e.g., porcine cells) with delayed emergence of cytosolic double-stranded RNA, the interferon response is late and viral dissemination is consequently enhanced.
Crystal Structures of GII.10 and GII.12 Norovirus Protruding Domains in Complex with Histo-Blood Group Antigens Reveal Details for a Potential Site of Vulnerability
Authors: Hansman Grant S. , Biertümpfel Christian , Georgiev Ivelin , McLellan Jason S. , Chen Lei , Zhou Tongqing , Katayama Kazuhiko , Kwong Peter D.
Published: April 2011

Noroviruses are the dominant cause of outbreaks of gastroenteritis worldwide, and interactions with human histo-blood group antigens (HBGAs) are thought to play a critical role in their entry mechanism. Structures of noroviruses from genogroups GI and GII in complex with HBGAs, however, reveal different modes of interaction. To gain insight into norovirus recognition of HBGAs, we determined crystal structures of norovirus protruding domains from two rarely detected GII genotypes, GII.10 and GII.12, alone and in complex with a panel of HBGAs, and analyzed structure-function implications related to conservation of the HBGA binding pocket. The GII.10- and GII.12-apo structures as well as the previously solved GII.4-apo structure resembled each other more closely than the GI.1-derived structure, and all three GII structures showed similar modes of HBGA recognition. The primary GII norovirus-HBGA interaction involved six hydrogen bonds between a terminal αfucose1-2 of the HBGAs and a dimeric capsid interface, which was composed of elements from two protruding subdomains. Norovirus interactions with other saccharide units of the HBGAs were variable and involved fewer hydrogen bonds. Sequence analysis revealed a site of GII norovirus sequence conservation to reside under the critical αfucose1-2 and to be one of the few patches of conserved residues on the outer virion-capsid surface. The site was smaller than that involved in full HBGA recognition, a consequence of variable recognition of peripheral saccharides. Despite this evasion tactic, the HBGA site of viral vulnerability may provide a viable target for small molecule- and antibody-mediated neutralization of GII norovirus.
Coxsackievirus B3-Induced Cellular Protrusions: Structural Characteristics and Functional Competence
Authors: Paloheimo Outi , Ihalainen Teemu O. , Tauriainen Sisko , Välilehto Outi , Kirjavainen Sanna , Niskanen Einari A. , Laakkonen Johanna P. , Hyöty Heikki , Vihinen-Ranta Maija
Published: April 2011

Virus-induced alterations in cell morphology play important roles in the viral life cycle. To examine the intracellular events of coxsackievirus B3 (CVB3) infection, green monkey kidney (GMK) cells were either inoculated with the virus or transfected with the viral RNA. Various microscopic and flow cytometric approaches demonstrated the emergence of CVB3 capsid proteins at 8 h posttransfection, followed by morphological transformation of the cells. The morphological changes included formation of membranous protrusions containing viral capsids, together with microtubules and actin. Translocation of viral capsids into these protrusions was sensitive to cytochalasin D, suggesting the importance of actin in the process. Three-dimensional (3D) live-cell imaging demonstrated frequent contacts between cellular protrusions and adjacent cells. Markedly, in spite of an increase in the cellular viral protein content starting 8 h postinfection, no significant decrease in cell viability or increase in the amount of early apoptotic markers was observed by flow cytometry by 28 h postinfection. Comicroinjection of viral RNA and fluorescent dextran in the presence of neutralizing virus antibody suggested that these protrusions mediated the spread of infection from one cell to another prior to virus-induced cell lysis. Altogether, the CVB3-induced cellular protrusions could function as a hitherto-unknown nonlytic mechanism of cell-to-cell transmission exploited by enteroviruses.
Ambivalent Role of the Innate Immune Response in Rabies Virus Pathogenesis
Authors: Chopy Damien , Pothlichet Julien , Lafage Mireille , Mégret Françoise , Fiette Laurence , Si-Tahar Mustapha , Lafon Monique
Published: April 2011

The neurotropic rabies virus (RABV) has developed several evasive strategies, including immunoevasion, to successfully infect the nervous system (NS) and trigger a fatal encephalomyelitis. Here we show that expression of LGP2, a protein known as either a positive or negative regulator of the RIG-I-mediated innate immune response, is restricted in the NS. We used a new transgenic mouse model (LGP2 TG) overexpressing LGP2 to impair the innate immune response to RABV and thus revealed the role of the RIG-I-mediated innate immune response in RABV pathogenesis. After infection, LGP2 TG mice exhibited reduced expression of inflammatory/chemoattractive molecules, beta interferon (IFN-β), and IFN-stimulated genes in their NS compared to wild-type (WT) mice, demonstrating the inhibitory function of LGP2 in the innate immune response to RABV. Surprisingly, LGP2 TG mice showed more viral clearance in the brain and lower morbidity than WT mice, indicating that the host innate immune response, paradoxically, favors RABV neuroinvasiveness and morbidity. LGP2 TG mice exhibited similar neutralizing antibodies and microglia activation to those of WT mice but showed a reduction of infiltrating CD4
+
T cells and less disappearance of infiltrating CD8
+
T cells. This occurred concomitantly with reduced neural expression of the IFN-inducible protein B7-H1, an immunoevasive protein involved in the elimination of infiltrated CD8
+
T cells. Our study shows that the host innate immune response favors the infiltration of T cells and, at the same time, promotes CD8
+
T cell elimination. Thus, to a certain extent, RABV exploits the innate immune response to develop its immunoevasive strategy.
Detection of Chronic Wasting Disease Prions in Salivary, Urinary, and Intestinal Tissues of Deer: Potential Mechanisms of Prion Shedding and Transmission
Authors: Haley Nicholas J. , Mathiason Candace K. , Carver Scott , Zabel Mark , Telling Glenn C. , Hoover Edward A.
Published: April 2011

Efficient horizontal transmission is a signature trait of chronic wasting disease (CWD) in cervids. Infectious prions shed into excreta appear to play a key role in this facile transmission, as has been demonstrated by bioassays of cervid and transgenic species and serial protein misfolding cyclic amplification (sPMCA). However, the source(s) of infectious prions in these body fluids has yet to be identified. In the present study, we analyzed tissues proximate to saliva, urine, and fecal production by sPMCA in an attempt to elucidate this unique aspect of CWD pathogenesis. Oropharyngeal, urogenital, and gastrointestinal tissues along with blood and obex from CWD-exposed cervids (comprising 27 animals and >350 individual samples) were analyzed and scored based on the apparent relative CWD burden. PrP
CWD
-generating activity was detected in a range of tissues and was highest in the salivary gland, urinary bladder, and distal intestinal tract. In the same assays, blood from the same animals and unseeded normal brain homogenate controls (
n
= 116 of 117) remained negative. The PrP-converting activity in peripheral tissues varied from 10
−11
- to 10
0
-fold of that found in brain of the same animal. Deer with highest levels of PrP
CWD
amplification in the brain had higher and more widely disseminated prion amplification in excretory tissues. Interestingly, PrP
CWD
was not demonstrable in these excretory tissues by conventional Western blotting, suggesting a low prion burden or the presence of protease-sensitive infectious prions destroyed by harsh proteolytic treatments. These findings offer unique insights into the transmission of CWD in particular and prion infection and trafficking overall.
Varicella-Zoster Virus Infection of Differentiated Human Neural Stem Cells
Authors: Pugazhenthi Subbiah , Nair Sreekala , Velmurugan Kalpana , Liang Qiaoling , Mahalingam Ravi , Cohrs Randall J. , Nagel Maria A. , Gilden Don
Published: April 2011

Primary varicella-zoster virus (VZV) infection in humans produces varicella (chickenpox), after which the virus becomes latent in ganglionic neurons. Analysis of the physical state of viral nucleic acid and virus gene expression during latency requires postmortem acquisition of fresh human ganglia. To provide an additional way to study the VZV-host relationship in neurons, we developed an
in vitro
model of infected differentiated human neural stem cells (NSCs). NSCs were induced to differentiate in culture dishes coated with poly-
l
-lysine and mouse laminin in the presence of fibroblast growth factor 2 (FGF-2), nerve growth factor (NGF), brain-derived neurotropic factor (BDNF), dibutyryl cyclic AMP, and retinoic acid. Immunostaining with neuronal (MAP2a and β-tubulin), astrocyte (GFAP), and oligodendrocyte (CNPase) markers revealed that differentiated neurons constituted approximately 90% of the cell population. These neurons were maintained in culture for up to 8 weeks. No cytopathic effect (CPE) developed in neurons infected with cell-free VZV (Zostavax vaccine) compared to human fetal lung fibroblasts infected with VZV. Weeks later, VZV DNA virus-specific transcripts (open reading frames [ORFs] 21, 29, 62, and 63) were detected in infected neurons, and dual immunofluorescence staining revealed the presence of VZV IE63 and gE exclusively in healthy-appearing neurons, but not in astrocytes. Neither the tissue culture medium nor a homogenate prepared from VZV-infected neurons produced a CPE in fibroblasts. VZV induced apoptosis in fibroblasts, as shown by activation of caspase 3 and by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining, but not in neurons. This model provides a unique
in vitro
system to study the VZV-neuronal relationship.
Multimerization of Adenovirus Serotype 3 Fiber Knob Domains Is Required for Efficient Binding of Virus to Desmoglein 2 and Subsequent Opening of Epithelial Junctions
Authors: Wang Hongjie , Li ZongYi , Yumul Roma , Lara Stephanie , Hemminki Akseli , Fender Pascal , Lieber André
Published: April 2011

Recently, we identified desmoglein 2 (DSG2) as the main receptor for a group of species B adenoviruses (Ads), including Ad3, a serotype that is widely distributed in the human population (H. Wang et al., Nat. Med.
17:
96–104, 2011). In this study, we have attempted to delineate structural details of the Ad3 interaction with DSG2. For CAR- and CD46-interacting Ad serotypes, attachment to cells can be completely blocked by an excess of recombinant fiber knob protein, while soluble Ad3 fiber knob only inefficiently blocks Ad3 infection. We found that the DSG2-interacting domain(s) within Ad3 is formed by several fiber knob domains that have to be in the spatial constellation that is present in viral particles. Based on this finding, we generated a small recombinant, self-dimerizing protein containing the Ad3 fiber knob (Ad3-K/S/Kn). Ad3-K/S/Kn bound to DSG2 with high affinity and blocked Ad3 infection. We demonstrated by confocal immunofluorescence and transmission electron microscopy analyses that Ad3-K/S/Kn, through its binding to DSG2, triggered the transient opening of intercellular junctions in epithelial cells. The pretreatment of epithelial cells with Ad3-K/S/Kn resulted in increased access to receptors that are localized in or masked by epithelial junctions, e.g., CAR or Her2/
neu
. Ad3-K/S/Kn treatment released CAR from tight junctions and thus increased the transduction of epithelial cells by a serotype Ad5-based vector. Furthermore, the pretreatment of Her2/
neu
-positive breast cancer cells with Ad3-K/S/Kn increased the killing of cancer cells by the Her2/
neu
-targeting monoclonal antibody trastuzumab (Herceptin). This study widens our understanding of how Ads achieve high avidity to their receptors and the infection of epithelial tissue. The small recombinant protein Ad3-K/S/Kn has practical implications for the therapy of epithelial cancer and gene/drug delivery to normal epithelial tissues.
Regulation of the Production of Infectious Genotype 1a Hepatitis C Virus by NS5A Domain III
Authors: Kim Seungtaek , Welsch Christoph , Yi MinKyung , Lemon Stanley M.
Published: April 2011

Although hepatitis C virus (HCV) assembly remains incompletely understood, recent studies with the genotype 2a JFH-1 strain suggest that it is dependent upon the phosphorylation of Ser residues near the C terminus of NS5A, a multifunctional nonstructural protein. Since genotype 1 viruses account for most HCV disease yet differ substantially in sequence from that of JFH-1, we studied the role of NS5A in the production of the H77S virus. While less efficient than JFH-1, genotype 1a H77S RNA produces infectious virus when transfected into permissive Huh-7 cells. The exchange of complete NS5A sequences between these viruses was highly detrimental to replication, while exchanges of the C-terminal domain III sequence (46% amino acid sequence identity) were well tolerated, with little effect on RNA synthesis. Surprisingly, the placement of the H77S domain III sequence into JFH-1 resulted in increased virus yields; conversely, H77S yields were reduced by the introduction of domain III from JFH-1. These changes in infectious virus yield correlated well with changes in the abundance of NS5A in RNA-transfected cells but not with RNA replication or core protein expression levels. Alanine replacement mutagenesis of selected Ser and Thr residues in the C-terminal domain III sequence revealed no single residue to be essential for infectious H77S virus production. However, virus production was eliminated by Ala substitutions at multiple residues and could be restored by phosphomimetic Asp substitutions at these sites. Thus, despite low overall sequence homology, the production of infectious virus is regulated similarly in JFH-1 and H77S viruses by a conserved function associated with a C-terminal Ser/Thr cluster in domain III of NS5A.
Temporal Variations in the Hepatitis C Virus Intrahost Population during Chronic Infection
Authors: Ramachandran Sumathi , Campo David S. , Dimitrova Zoya E. , Xia Guo-liang , Purdy Michael A. , Khudyakov Yury E.
Published: April 2011

The intrahost evolution of hepatitis C virus (HCV) holds keys to understanding mechanisms responsible for the establishment of chronic infections and to development of a vaccine and therapeutics. In this study, intrahost variants of two variable HCV genomic regions, HVR1 and NS5A, were sequenced from four treatment-naïve chronically infected patients who were followed up from the acute stage of infection for 9 to 18 years. Median-joining network analysis indicated that the majority of the HCV intrahost variants were observed only at certain time points, but some variants were detectable at more than one time point. In all patients, these variants were found organized into communities or subpopulations. We hypothesize that HCV intrahost evolution is defined by two processes: incremental changes within communities through random mutation and alternations between coexisting communities. The HCV population was observed to incrementally evolve within a single community during approximately the first 3 years of infection, followed by dispersion into several subpopulations. Two patients demonstrated this pattern of dispersion for the rest of the observation period, while HCV variants in the other two patients converged into another single subpopulation after ∼9 to 12 years of dispersion. The final subpopulation in these two patients was under purifying selection. Intrahost HCV evolution in all four patients was characterized by a consistent increase in negative selection over time, suggesting the increasing HCV adaptation to the host late in infection. The data suggest specific staging of HCV intrahost evolution.
Proteomic Characterization of Pseudorabies Virus Extracellular Virions
Authors: Kramer T. , Greco T. M. , Enquist L. W. , Cristea I. M.
Published: April 2011

Pseudorabies virus (PRV), a member of the
Alphaherpesvirinae
, has a complex multilayered extracellular virion that is structurally conserved among other herpesviruses. PRV virions contain a double-stranded DNA genome within a proteinaceous capsid surrounded by the tegument, a layer of viral and cellular proteins. The envelope layer, which encloses the capsid and tegument, contains viral transmembrane proteins anchored in a phospholipid bilayer. The viral and host proteins contained within virions execute important functions during viral spread and pathogenesis, but a detailed understanding of the composition of PRV virions has been lacking. In this report, we present the first comprehensive proteomic characterization of purified PRV virions by mass spectrometry using two complementary approaches. To exclude proteins present in the extracellular medium that may nonspecifically associate with virions, we also analyzed virions treated with proteinase K and samples prepared from mock-infected cells. Overall, we identified 47 viral proteins associated with PRV virions, 40 of which were previously localized to the capsid, tegument, and envelope layers using traditional biochemical approaches. Additionally, we identified seven viral proteins that were previously undetected in virions, including pUL8, pUL20, pUL32, pUL40 (RR2), pUL42, pUL50 (dUTPase), and Rsp40/ICP22. Furthermore, although we did not enrich for posttranslational modifications, we detected phosphorylation of four virion proteins: pUL26, pUL36, pUL46, and pUL48. Finally, we identified 48 host proteins associated with PRV virions, many of which have known functions in important cellular pathways such as intracellular signaling, mRNA translation and processing, cytoskeletal dynamics, and membrane organization. This analysis extends previous work aimed at determining the composition of herpesvirus virions and provides novel insights critical for understanding the mechanisms underlying PRV entry, assembly, egress, spread, and pathogenesis.
Bovine mycoplasmosis: silent and deadly
Authors: Nicholas R. A. J.
Published: April 2011

The first international conference on bovine mycoplasmosis, held in Saskatoon, Canada, in 2009, marked an official recognition of the importance of mycoplasmas in bovine respiratory disease, mastitis, arthritis and other disorders. Furthermore, bovine mycoplasmosis has been identified as a major emerging infectious disease by the European Community 7th Framework Programme under the EMIDA ERA‐NET, which is funding a three‐year study into the development of improved diagnosis and control of bovine mycoplasmosis involving the UK, Germany, Italy and Israel. These developments coincide with the publication of the complete sequence of the Mycoplasma bovis genome, which offers fresh insight into the properties of this tiny wall‐less bacterium. With this in mind, it is an appropriate time to review our knowledge of cattle diseases caused by mycoplasmas.
ON THE GOUSSAROV–POLYAK–VIRO FINITE-TYPE INVARIANTS AND THE VIRTUALIZATION MOVE
Authors: CHRISMAN MICAH W.
Published: April 2011

In this paper, it is shown that there are no nonconstant Goussarov–Polyak–Viro finite-type invariants that are invariant under the virtualization move. As an immediate corollary, we obtain the theorem of [1] which states none of the Birman coefficients of the Jones–Kauffman polynomial are of GPV finite type.
ALEXANDER–LIN TWISTED POLYNOMIALS
Authors: SILVER DANIEL S. , WILLIAMS SUSAN G.
Published: April 2011

Lin's definition of twisted Alexander knot polynomial is extended for finitely presented groups. Cha's fibering obstruction theorem is generalized. The group of the 2-twist-spun trefoil knot is seen to have a faithful representation that yields a trivial twisted Alexander polynomial.
INVARIANTS OF LINKS IN 3-MANIFOLDS AND SPLITTING PROBLEM OF TEXTILE STRUCTURES
Authors: GRISHANOV S. A. , VASSILIEV V. A.
Published: April 2011

An infinite family of invariants of multicomponent links in 3-manifolds is introduced and used to prove the non-splitting and non-equivalence of textile structures.
ON THE STRUCTURE AND AUTOMORPHISM GROUP OF FINITE ALEXANDER QUANDLES
Authors: FERMAN AMIEL , NOWIK TAHL , TEICHER MINA
Published: April 2011

We prove that an Alexander quandle of prime order is generated by any pair of distinct elements. Furthermore, we prove for such a quandle that any ordered pair of distinct elements can be sent to any other such pair by an automorphism of the quandle.
STRATEGIC ENVIRONMENTAL ASSESSMENT AS A TOOL TO CONTRIBUTE TO HIGH-LEVEL POLICY OBJECTIVES
Authors: JOÃO ELSA , MCLAUCHLAN ANNA
Published: April 2011

Strategic environmental assessment (SEA) has often been identified as a key tool to contribute to sustainable development. This special issue of the Journal of Environmental Assessment Policy and Management focuses on European SEA practice between 2003 and 2010 to critically evaluate SEA links to sustainable development and similar, difficult to define, high-level policy objectives: democratisation, good governance, agri-environmental objectives, and environmental justice. The papers centre upon the three main topics covered by the different research: SEA outcomes being directly related to policy goals; an analysis of the absence of SEA applied to "positive" policy objectives; and the realism of associating SEA with high-level policy objectives. In particular, this paper calls for greater critical engagement with this latter topic, identifying a need to examine why associations are made between SEA and policy goals.
STRATEGIC ENVIRONMENTAL ASSESSMENT AND THE DEMOCRATISATION OF SPATIAL PLANNING
Authors: BONIFAZI ALESSANDRO , REGA CARLO , GAZZOLA PAOLA
Published: April 2011

This paper investigates the potential of Strategic Environmental Assessment (SEA) to contribute to the high-level policy objective of fostering democratisation in spatial planning. Democratic SEA is conceptualised by discussing environmental citizenship, public control over policy making, and participants' empowerment. The empirical investigation is based on the analysis of SEA documents produced during 25 municipal spatial planning processes in Italy between 2004 and 2010. The study found that advances are more evident in the creation of cross-sectoral governance networks than in the involvement of citizens and civil-society organisations. SEA seems to be increasing transparency and expanding the scope of democratic control over spatial planning decisions, though the boundaries between experts, decision makers and citizens are clearly demarcated. To strengthen democratisation processes, the paper argues that all participants should consider themselves equally responsible within SEA networks, and be ready to question alternative environmental value systems that underpin spatial planning processes.
EFFECTIVENESS OF POLICY LEVEL ENVIRONMENTAL AND SUSTAINABILITY ASSESSMENT: CHALLENGES AND LESSONS FROM RECENT PRACTICE
Authors: EALES RICHARD P. , SHEATE WILLIAM R.
Published: April 2011

This paper explores the challenges and lessons from recent practice and experience of applying tools such as Strategic Environmental Assessment (SEA) and Sustainability Appraisal at the policy level in the UK and Europe. It investigates whether or not these tools have been effective and helped to deliver more sustainable development at the high level of national policy development. The analysis is illustrated by case examples from the UK, such as Eco-towns and Energy Planning. The paper concludes that the current performance by the UK Government in implementing the SEA Directive for national level strategic actions is far from exemplary. At the root of the problem is the poor consideration and evaluation of reasonable alternatives, the fundamentally weak conception of sustainability adopted and the apparent perception that having to undertake an assessment and comply with the SEA Directive is a hurdle, rather than a useful mechanism for helping to deliver better and more sustainable evidence-based policy making.
GOVERNMENTALITY AND STRATEGIC ENVIRONMENTAL ASSESSMENT: CHALLENGING THE SEA/GOOD GOVERNANCE NEXUS
Authors: SCOTT CAROLINE
Published: April 2011

Strategic Environmental Assessment (SEA) has been associated with "good governance" by bodies at national and international levels including the World Bank, OECD, and UK and Scottish Governments. Typically involving components such as transparency, accountability, public participation and partnership working, this SEA/good governance nexus has been promoted in Scotland where the government sees SEA as central to its sustainable development aspirations. Using a governmentality lens to view SEA as a technique seeking to instil environmentally-focused governance, the paper examines the operation of the SEA/good governance nexus in the SEA process of one Scottish case study, a road corridor development framework undertaken between 2006 and 2008. The paper exposes instances of resistance to both the democratising elements of good governance and to SEA itself as the public and statutory Consultation Authorities find their efforts to constructively engage with the SEA process thwarted. This reveals that, in the case studied, the SEA/good governance nexus, as a high-level policy objective, is more aspiration than reality.
NONINVASIVE PROBING OF THE NEUROVASCULAR SYSTEM IN HUMAN BONE/BONE MARROW USING NEAR-INFRARED LIGHT
Authors: BINZONI TIZIANO , VAN DE VILLE DIMITRI
Published: April 2011

Understanding the mechanisms of interaction between bone/bone marrow, circulatory system and nervous system is of great interest due to the potential clinical impact. In humans, the amount of knowledge in this domain remains relatively limited due to the extreme difficulty to monitor these tissues continuously, noninvasively and for long or repeated periods of time. A typical difficult task would be, for example, to continuously monitor bone/bone marrow blood perfusion, hemoglobin oxygen saturation or blood volume and study their dependence on the activity of the autonomic nervous system. In this review article, we want to show that near-infrared light might be utilized to solve these problems in part. We hope that the present analysis will stimulate future studies in this domain, for which near-infrared light appears as the best available technology today.
OPTICAL MEASUREMENT OF PHOTOSENSITIZER CONCENTRATION IN VIVO
Authors: AUSTWICK MARTIN R. , WOODHAMS JOSEPHINE H. , CHALAU VADZIM , MOSSE CHARLES A. , ELIOT CAROLINE , LOVAT LAURENCE , MACROBERT ALEXANDER J. , BIGIO IRVING J. , BOWN STEPHEN G.
Published: April 2011

Most techniques for measuring tissue concentrations of drugs are invasive, time-consuming, and often require the removal of tissue or body fluids. Optical pharmacokinetics (OP) is a minimally invasive alternative giving an immediate result. Pulses of white light are directed at the tissue of interest using a fiber optic probe. Scattered light is detected by a second fiber immediately adjacent to the first in the same probe (separation 1.7 mm). Using the photosensitizer disulfonated aluminium phthalocyanine ( AlS2Pc ), OP measurements were made in phantoms and on the mouth, stomach, colon, skin, and liver of normal rats 1 and 24 h after intravenous AlS2Pc administration. AlS2Pc concentration was determined by calculating the area under the curve (AUC) in the spectral region around the peak drug absorption or measuring the height of the peak. Spectral baseline interpolation removed the need for pre-drug, control optical measurements. OP measurements correlated well with values from alkali chemical extraction (CE) of the corresponding tissues, (R2 0.87–0.97). OP measurements in the mouth also correlated with CE of less accessible internal organs (R2 0.77–0.88). In phantoms, the lowest detectable concentration was 0.1 μg/g. In vivo, results were limited by the lower accuracy in the CE measurements but were almost certainly comparable. An incidental finding was a 12–15 nm red shifted component in the spectra observed 1 h after drug administration, suggesting partitioning of the drug in different microenvironment compartments, which could prove to be of considerable interest in future studies. In conclusion, OP shows promise for real-time measurement of concentrations of drugs with suitable absorption peaks.
Role of Rac1 in regulation of NOX5-S function in Barrett's esophageal adenocarcinoma cells
Authors: Hong Jie , Resnick Murray , Behar Jose , Wands Jack , DeLellis Ronald A. , Cao Weibiao
Published: April 2011

We have shown that a novel NADPH oxidase isoform, NOX5-S, is the major isoform of NADPH oxidases in an esophageal adenocarcinoma (EA) cell line, FLO, and is overexpressed in Barrett's mucosa with high-grade dysplasia. NOX5-S is responsible for acid-induced reactive oxygen species production. In this study, we found that mRNA levels of NOX5-S were significantly higher in FLO EA cells than in the normal human esophageal squamous cell line HET-1A or in a Barrett cell line, BAR-T. The mRNA levels of NOX5-S were also significantly increased in EA tissues. The data suggest that NOX5-S may be important in the development of EA. Mechanisms of functional regulation of NOX5-S are not fully understood. We show that small G protein Rac1 was present in HET-1A cells, BAR-T cells, and EA cell lines FLO and OE33. Rac1 protein levels were significantly higher in FLO and OE33 cells than in HET-1A or BAR-T cells. Knockdown of Rac1 with Rac1 small interfering RNA significantly decreased acid-induced increase in H2O2 production in FLO EA cells. Overexpression of constitutively active Rac1 significantly increased H2O2 production, an increase that was blocked by knockdown of NOX5-S. By immunofluorescence staining and immunoprecipitation, we found that NOX5-S was present in the cytosol of FLO EA cells and colocalized with Rac1 and SERCA1/2 Ca2+-ATPase which is located in the endoplasmic reticulum membrane. We conclude that Rac1 may be important in activation of NOX5-S in FLO EA cells.