bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibitors of Respiratory Syncytial Virus Replication Target Cotranscriptional mRNA Guanylylation by Viral RNA-Dependent RNA Polymerase
Authors: Liuzzi Michel , Mason Stephen W. , Cartier Mireille , Lawetz Carol , McCollum Robert S. , Dansereau Nathalie , Bolger Gordon , Lapeyre Nicole , Gaudette Yvon , Lagacé Lisette , Massariol Marie-Josée , Dô Florence , Whitehead Paul , Lamarre Lyne , Scouten Erika , Bordeleau Josée , Landry Serge , Rancourt Jean , Fazal Gulrez , Simoneau Bruno 
Published: September 2005

ABSTRACT
Respiratory
syncytial virus (RSV) is a major cause of respiratory illness in
infants, immunocompromised patients, and the elderly. New antiviral
agents would be important tools in the treatment of acute RSV disease.
RSV encodes its own RNA-dependent RNA polymerase that is responsible
for the synthesis of both genomic RNA and subgenomic mRNAs. The viral
polymerase also cotranscriptionally caps and polyadenylates the RSV
mRNAs at their 5′ and 3′ ends, respectively. We have
previously reported the discovery of the first nonnucleoside
transcriptase inhibitor of RSV polymerase through high-throughput
screening. Here we report the design of inhibitors that have improved
potency both in vitro and in antiviral assays and that also exhibit
activity in a mouse model of RSV infection. We have isolated virus with
reduced susceptibility to this class of inhibitors. The mutations
conferring resistance mapped to a novel motif within the RSV L gene,
which encodes the catalytic subunit of RSV polymerase. This motif is
distinct from the catalytic region of the L protein and bears some
similarity to the nucleotide binding domain within nucleoside
diphosphate kinases. These findings lead to the hypothesis that this
class of inhibitors may block synthesis of RSV mRNAs by inhibiting
guanylylation of viral transcripts. We show that short transcripts
produced in the presence of inhibitor in vitro do not contain a
5′ cap but, instead, are triphosphorylated, confirming this
hypothesis. These inhibitors constitute useful tools for elucidating
the molecular mechanism of RSV capping and represent valid leads for
the development of novel anti-RSV
therapeutics.
Hepatocyte-Like Cells Transdifferentiated from a Pancreatic Origin Can Support Replication of Hepatitis B Virus
Authors: Wang Robert Yung-Liang , Shen Chia-Ning , Lin Min-Hui , Tosh David , Shih Chiaho 
Published: September 2005

ABSTRACTRecently, a rat pancreatic cell line (AR42J-B13) was shown to transdifferentiate to hepatocyte-like cells upon induction with dexamethasone (Dex). The aim of this study is to determine whether transdifferentiated hepatocytes can indeed function like bona fide liver cells and support replication of hepatotropic hepatitis B virus (HBV). We stably transfected AR42J-B13 cells with HBV DNA and examined the expression of hepatocyte markers and viral activities in control and transdifferentiated cells. A full spectrum of HBV replicative intermediates, including covalently closed circular DNA (cccDNA) and Dane particles, were detected only after induction with Dex and oncostatin M. Strikingly, the small envelope protein and RNA of HBV were increased by 40- to 100-fold upon induction. When HBV RNAs were examined by primer extension analysis, novel core- and precore-specific transcripts were induced by Dex which initiated at nucleotide (nt) 1820 and nt 1789, respectively. Most surprisingly, another species of core-specific RNA, which initiates at nt 1825, is always present at almost equal intensity before and after Dex treatment, a result consistent with Northern blot analysis. The fact that HBV core protein is dramatically produced only after transdifferentiation suggests the possibility of both transcriptional and translational regulation of HBV core antigen in HBV-transfected AR42J-B13 cells. Upon withdrawal of Dex, HBV replication and gene expression decreased rapidly—less than 50% of the cccDNA remained detectable in 1.5 days. Our studies demonstrate that the transdifferentiated AR42J-B13 cells can function like bona fide hepatocytes. This system offers a new opportunity for basic research of virus-host interactions and pancreatic transdifferentiation.
SAT: a Late NS Protein of Porcine Parvovirus
Authors: Zádori Zoltán , Szelei József , Tijssen Peter 
Published: September 2005

ABSTRACT

The genomes of all members of the
Parvovirus
genus were found to contain a small open reading frame (ORF), designated SAT, with a start codon four or seven nucleotides downstream of the VP2 initiation codon. Green fluorescent protein or FLAG fusion constructs of SAT demonstrated that these ORFs were expressed. Although the SAT proteins of the different parvoviruses are not particularly conserved, they were all predicted to contain a membrane-spanning helix, and mutations in this hydrophobic stretch affected the localization of the SAT protein. SAT colocalized with calreticulin in the membranes of the endoplasmic reticulum and the nucleus. A knockout mutant (SAT
−
), with an unmodified VP sequence, showed a “slow-spreading” phenotype. These knockout mutants could be complemented with VP2
−
SAT
+
mutant. The SAT protein is a late nonstructural (NS) protein, in contrast to previously identified NS proteins, since it is expressed from the same mRNA as VP2.
Human Papillomavirus Type 16 E1
∧
E4 Contributes to Multiple Facets of the Papillomavirus Life Cycle
Authors: Nakahara Tomomi , Peh Woei Ling , Doorbar John , Lee Denis , Lambert Paul F. 
Published: September 2005

ABSTRACT

The life cycle of human papillomaviruses (HPVs) is tightly linked to the differentiation program of the host's stratified epithelia that it infects. E1
∧
E4 is a viral protein that has been ascribed multiple biochemical properties of potential biological relevance to the viral life cycle. To identify the role(s) of the viral E1
∧
E4 protein in the HPV life cycle, we characterized the properties of HPV type 16 (HPV16) genomes harboring mutations in the E4 gene in NIKS cells, a spontaneously immortalized keratinocyte cell line that when grown in organotypic raft cultures supports the HPV life cycle. We learned that E1
∧
E4 contributes to the replication of the viral plasmid genome as a nuclear plasmid in basal cells, in which we also found E1
∧
E4 protein to be expressed at low levels. In the suprabasal compartment of organotypic raft cultures harboring E1
∧
E4 mutant HPV16 genomes there were alterations in the frequency of suprabasal cells supporting DNA synthesis, the levels of viral DNA amplification, and the degree to which the virus perturbs differentiation. Interestingly, the comparison of the phenotypes of various mutations in E4 indicated that the E1
∧
E4 protein-encoding requirements for these various processes differed. These data support the hypothesis that E1
∧
E4 is a multifunctional protein and that the different properties of E1
∧
E4 contribute to different processes in both the early and late stages of the virus life cycle.
Homotypic Interaction of Bunyamwera Virus Nucleocapsid Protein
Authors: Leonard Vincent H. J. , Kohl Alain , Osborne Jane C. , McLees Angela , Elliott Richard M. 
Published: September 2005

ABSTRACT
The bunyavirus nucleocapsid protein, N, plays a central role in viral replication in encapsidating the three genomic RNA segments to form functional templates for transcription and replication by the viral RNA-dependent RNA polymerase. Here we report functional mapping of interacting domains of the Bunyamwera orthobunyavirus N protein by yeast and mammalian two-hybrid systems, immunoprecipitation experiments, and chemical cross-linking studies. N forms a range of multimers from dimers to high-molecular-weight structures, independently of the presence of RNA. Deletion of the N- or C-terminal domains resulted in loss of activity in a minireplicon assay and a decreased capacity for N to form higher multimers. Our data suggest a head-to-head and tail-to-tail multimerization model for the orthobunyavirus N protein.
A Role for Herpesvirus Entry Mediator as the Receptor for Herpes Simplex Virus 1 Entry into Primary Human Trabecular Meshwork Cells
Authors: Tiwari Vaibhav , Clement Christian , Scanlan Perry M. , Kowlessur Devanand , Yue Beatrice Y. J. T. , Shukla Deepak 
Published: September 2005

ABSTRACTThe human eye is an important target for infection with herpes simplex virus 1 (HSV-1). Damage to cells forming the trabeculum of the eye by HSV-1 infection could contribute to the development of glaucoma, a major blinding disease. Primary cultures of human trabecular meshwork cells were used as an in vitro model to demonstrate the ability of HSV-1 to enter into and establish a productive infection of the trabeculum. Blocking of entry by anti-herpesvirus entry mediator (HVEM) antibody implicated HVEM as the major receptor for HSV-1 infection.
Kaposi's Sarcoma-Associated Herpesvirus Virion-Induced Transcription Activation of the ORF50 Immediate-Early Promoter
Authors: Lu Fang , Day Latasha , Lieberman Paul M. 
Published: September 2005

ABSTRACT

Lytic cycle reactivation of Kaposi's sarcoma-associated herpesvirus (KSHV) can be initiated by transcription activation of the ORF50 immediate-early (IE) gene promoter (ORF50p). We provide evidence that KSHV virions stimulate transcription of ORF50p. Virion activation was resistant to UV inactivation and cycloheximide treatment. The virion-responsive element was mapped to core promoter region −150 to + 1 relative to the ORF50 initiation codon. Electrophoretic mobility shift assays and chromatin immunoprecipitation suggest that KSHV virions indirectly alter the protein composition and chromatin modifications at ORF50p. These data suggest that KSHV virions possess an IE
trans
-inducing function similar to that observed in alpha- and betaherpesviruses.
Cytogenetic differences in breast cancer samples between German and Japanese patients
Authors: Packeisen J , Nakachi K , Boecker W , Brandt B , Buerger H 
Published: September 2005

Background:Japanese and German breast cancer cases differ substantially in the frequency ofegframplification.Aims:To unravel further the cytogenetic differences between Japanese and German breast cancer cases.Methods:Forty one Japanese breast cancer cases were evaluated by means of comparative genomic hybridisation (CGH). The results were compared with the CGH results from 161 German breast cancer cases.Results:The mean number of genetic alterations/case was significantly higher in German premenopausal patients with breast cancer than in their Japanese counterparts. Japanese breast cancer cases revealed a higher number of chromosome 17p losses. Losses of 8p were associated with oestrogen receptor (ER) negativity in Japanese patients with breast cancer, whereas in the German patients gains of 3q and 6q were associated with the lack of ER expression.Conclusions:The interethnic differences of invasive breast cancer are reflected by cytogenetic aberrations, which are also associated with the differential expression of the ER.
Dominantly inherited β thalassaemia intermedia caused by a new single nucleotide deletion in exon 2 of the β globin gene: Hb morgantown (β91 CTG>CG)
Authors: Luo H-Y , Tang W , Eung S H , Coad J E , Canfield P , Keller F , Crowell E H , Steinberg M H , Chui D H K 
Published: September 2005

Family members in multiple generations of an Irish–American family were investigated for moderate to severe microcytic anaemia, inherited in an autosomal dominant fashion. A novel frameshift mutation of the β globin gene was discovered. This study highlights the importance of considering dominantly inherited β thalassemia in the investigation of anaemia, even in patients with ethnic backgrounds not usually associated with β thalassaemia.
Suppressor of cytokine signalling 2 (SOCS-2) expression in breast carcinoma
Authors: Farabegoli F , Ceccarelli C , Santini D , Taffurelli M 
Published: September 2005

Aims:To investigate SOCS-2 (suppressor of cytokine signalling 2) protein expression in breast carcinoma samples in relation to biopathological parameters and survival.Methods:A polyclonal antibody against SOCS-2 was used to study 50 archival breast carcinoma samples, collected from 1993 to 1995. The presence of SOCS-2 protein was investigated in relation to clinical and biological parameters used in breast cancer pathology. Fluorescence in situ hybridisation (FISH) was used to study whether SOCS-2 expression was related to SOCS-2 gene copy number.Results:SOCS-2 protein was expressed in 34 of 50 breast carcinoma samples and was positively associated with low grade, low nuclear grade, and p27 protein. SOCS-2 expression was inversely related to Ki-67, cyclin A, retinoblastoma protein (pRb), and the epidermal growth factor receptor (EGFR). No relation with overall survival was demonstrated. SOCS-2 amplification was found in three samples. No relation between the number of FISH signals and SOCS-2 expression was found.Conclusions:The significant correlation seen between SOCS-2 expression, grade, nuclear grade, p27, Ki-67, cyclin A, pRb, and EGFR labelling strongly supports the hypothesis that SOCS-2 loss might be related to cell proliferation and tumour growth in breast carcinoma. Gene copy number changes did not seem to play a role in SOCS-2 regulation and expression; other mechanisms might be involved and deserve further study.
A shift to a peripheral Th2-type cytokine pattern during the carcinogenesis of cervical cancer becomes manifest in CIN III lesions
Authors: Bais A G , Beckmann I , Lindemans J , Ewing P C , Meijer C J L M , Snijders P J F , Helmerhorst T J M 
Published: September 2005

Background:A shifted balance between T helper 1 (Th1)-type and Th2-type cytokines has been hypothesised in cervical dysplasiaAims:To evaluate possible deregulation of the cytokine network by estimating the expression of peripheral cytokines in different stages of cervical disease and in relation to the presence or absence of high risk human papillomavirus (HR-HPV).Methods:Twenty one HR-HPV positive women with high grade cervical intraepithelial neoplasia (CIN II–III) and 12 patients with invasive cervical carcinoma formed the study groups. Two control groups consisted of 10 HR-HPV positive and 11 HR-HPV negative women without CIN. Differences in leucocyte subgroups were evaluated by a differential leucocyte count. Plasma concentrations of tumour necrosis factor α (TNFα), TNFα receptors TNFRI and TNFRII, interferon γ (IFNγ), interleukin 2 (IL-2), IL-12, IL-4, and IL-10 were determined by enzyme linked immunosorbent assays.Results:Leucocyte counts in patients with CIN III and carcinoma were significantly higher than in controls. Plasma IFNγ concentrations were significantly lower in patients with CIN III and carcinoma than in women with CIN II or controls. Plasma concentrations of IL-12, IL-2, IL-4, and TNFα did not differ significantly between groups, but significantly lower plasma concentrations of TNFRII were found in CIN III and carcinoma compared with CIN II. IL-10 was detected with increased frequency in the plasma of patients with CIN III and carcinoma.Conclusions:These results indicate that a shift to a Th2-type cytokine pattern during the carcinogenesis of cervical cancer occurs in women with CIN III lesions.
Gene amplification and protein expression of EGFR and HER2 by chromogenic in situ hybridisation and immunohistochemistry in atypical adenomatous hyperplasia and adenocarcinoma of the lung
Authors: Awaya H , Takeshima Y , Furonaka O , Kohno N , Inai K 
Published: September 2005

Aims:To investigate the importance of gene amplification and EGFR (epidermal growth factor receptor) and HER2 protein expression during the progression of adenocarcinoma of the lung.Methods:EGFR and HER2 gene amplification was examined in atypical adenomatous hyperplasia (AAH), bronchioloalveolar carcinoma (BAC), and adenocarcinoma with mixed subtypes (MX) by chromogenic in situ hybridisation (CISH), and protein expression was examined by immunohistochemistry using paraffin wax embedded tissues.Results:EGFR and HER2 gene amplification was found in four and two of 86 cases, respectively, and was detected only in the invasive components of MX. EGFR and HER2 protein expression was seen in 24 and 18 of 86 cases, respectively. EGFR and HER2 proteins were not expressed in AAH but were expressed in one BAC case each. EGFR and HER2 proteins were expressed in 23 and 17 of 55 adenocarcinomas with MX. EGFR and HER2 protein expression was seen more often in the invasive components than in the BAC components of MX, and increased significantly as lesions progressed from AAH to BAC, early MX, and overt MX. Because EGFR and HER2 protein expression was frequently seen without gene amplification, other mechanisms apart from gene amplification may be associated with protein expression.Conclusions:EGFR and HER2 gene amplification may be a late event and EGFR and HER2 protein expression may be associated with the development of adenocarcinoma of the lung.
The influence of extent of surgical margin positivity on prostate specific antigen recurrence
Authors: Emerson R E , Koch M O , Jones T D , Daggy J K , Juliar B E , Cheng L 
Published: September 2005

Background:Positive surgical margins are an adverse prognostic factor in patients undergoing prostatectomy for prostate cancer. The extent of margin positivity varies and its influence on clinical outcome is uncertain.Aims:To evaluate the linear extent of margin positivity and the number and location of positive sites as prognostic indicators in a series of prostatectomy specimens evaluated with the whole mount technique.Methods:Eighty six consecutive margin positive prostatectomy specimens were evaluated, and all pathology data were collected prospectively. The linear extent of margin positivity was measured with an ocular micrometer and the total extent of all positive sites was summed. The total number of sites with positive margins and anatomical sites of the positive margins were analysed.Results:The linear extent of margin positivity ranged from 0.01 to 68 mm (mean, 6.8; median, 3.0) and was associated with prostate specific antigen (PSA) recurrence in univariate logistic regression (p = 0.031). In addition, the extent of margin positivity weakly correlated with preoperative PSA (p = 0.017) and tumour volume (p = 0.013), but not with age, prostate weight, Gleason score, pathological stage, or perineural invasion. The total number of positive sites was significantly higher in patients with PSA recurrence (p = 0.037). The location of the positive margin site was not associated with PSA recurrence. The extent of margin positivity correlated with PSA recurrence in univariate analysis, although it had only marginal predictive value when adjusted for Gleason score (p = 0.076).Conclusions:The extent of margin positivity correlates with PSA recurrence in univariate analysis, although it has no predictive value independent of Gleason score.
Molecular pathology of ataxia telangiectasia
Authors: Taylor A M R , Byrd P J 
Published: September 2005

Ataxia telangiectasia (A-T) is one of a group of autosomal recessive cerebellar ataxias. Presentation is usually by the age of 2 years and ataxia of both upper and lower limbs develops, such that by early teenage most patients require a wheelchair for mobility. Speech and eye movement are also affected. Other important features are t(7;14) translocations, immunodeficiency, a high serum α fetoprotein concentration, growth retardation, telangiectasia—most noticeably on the bulbar conjunctiva—and a very high risk of developing a lymphoid tumour. Patients also show an increased sensitivity to ionising radiation. The classic form of A-T results from the presence of two truncatingATMmutations, leading to total loss of the ATM protein, a protein kinase. Importantly, A-T shows clinical heterogeneity, including milder forms where neurological progression may be slower or of later onset. In these cases there is a correlation between the preservation of neurological function, decreased radiosensitivity, and the degree of retained ATM protein kinase activity. Considerable scope remains for understanding the progress of the disorder in relation to the types ofATMmutation present.
Marsh stars in Liverpool
Authors: Agarwal A , Helliwell T R 
Published: September 2005

Introduction/Methods:After the recognition of contaminating algae on histopathological sections stained by periodic acid Schiff (PAS) and Grocott methods, a detailed audit was undertaken to assess the extent of contamination and its possible source.Results:The contaminating organism was a member of the staurastrum genus of Chlorophyta, star shaped organisms commonly found in fresh water marshes. The organisms were seen on sections stained by the diastase-PAS, PAS, or Grocott methods and on cytological preparations between July 2003 and May 2004. It is unlikely that contamination of water baths or concentrated staining solutions was to blame, and a more general contamination of the laboratory water supply is the most likely source.Conclusions:Contaminating organisms may appear on histological and cytological material and their nature and source should be investigated. Although in this instance, confusion with pathologically important organisms was minimal, algae may occasionally cause significant disease.
KSHV/HHV-8 associated lymph node based lymphomas in HIV seronegative subjects. Report of two cases with anaplastic large cell morphology and plasmablastic immunophenotype
Authors: Carbone A , Gloghini A , Vaccher E , Marchetti G , Gaidano G , Tirelli U 
Published: September 2005

Background:Kaposi sarcoma associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) associated lymphomas, which often develop in human immunodeficiency virus (HIV) infected patients with advanced AIDS, present predominantly as primary effusion lymphoma (PEL) or, less frequently, as “solid” extracavitary based lymphomas, associated with serous effusions. These last lymphomas, also called “solid PEL”, have been reported before the development of an effusion lymphoma and after resolution of PEL. Interestingly, KSHV/HHV-8 associated lymphomas that present as solid or extracavitary based lesions in HIV seropositive patients without serous effusions have been reported recently.Methods/Results:This paper provides evidence for the existence of a previously undescribed KSHV/HHV-8 associated lymphoma in HIV seronegative patients without serous effusions. These lymphomas exhibit a predilection for the lymph nodes and display anaplastic large cell morphology. These tumours were completely devoid of common cell type specific antigens, including epithelial and melanocytic cell markers. B and T cell associated antigens and other commonly used lymphoid markers were absent or weakly demonstrable in a fraction of the tumour cells. Conversely, immunohistochemical studies showed strong immunostaining with plasma cell reactive antibodies.Conclusions:Analysis of viral infection and immunohistological studies are of primary importance to define this lymph node based KSHV/HHV-8 associated lymphoma with anaplastic large cell morphology and plasmablastic immunophenotype occurring in HIV seronegative patients without serous effusions.