Science.gov

Sample records for human polyomavirus workshop

  1. Human polyomavirus related to African green monkey lymphotropic polyomavirus.

    PubMed

    Sauvage, Virginie; Foulongne, Vincent; Cheval, Justine; Ar Gouilh, Meriadeg; Pariente, Kevin; Dereure, Olivier; Manuguerra, Jean Claude; Richardson, Jennifer; Lecuit, Marc; Burguière, Ana; Caro, Valérie; Eloit, Marc

    2011-08-01

    While studying the virome of the skin surface of a patient with a Merkel cell carcinoma (MCC) by using unbiased, high-throughput sequencing, we identified a human polyomavirus nearly identical to human polyomavirus 9, a virus recently reported in blood and urine of renal transplantion patients and closely related to the African green monkey lymphotropic polyomavirus. Specific PCR analysis further identified this virus in 2/8 patients with MCC but in only 1/111 controls without MCC. This virus was shed for ≥20 months by the MCC index patient and was on the skin of the spouse of the index patient. These results provide information on the viral ecology of human skin and raise new questions regarding the pathology of virus-associated skin disorders.

  2. The role of Merkel cell polyomavirus and other human polyomaviruses in emerging hallmarks of cancer.

    PubMed

    Moens, Ugo; Rasheed, Kashif; Abdulsalam, Ibrahim; Sveinbjørnsson, Baldur

    2015-04-10

    Polyomaviruses are non-enveloped, dsDNA viruses that are common in mammals, including humans. All polyomaviruses encode the large T-antigen and small t-antigen proteins that share conserved functional domains, comprising binding motifs for the tumor suppressors pRb and p53, and for protein phosphatase 2A, respectively. At present, 13 different human polyomaviruses are known, and for some of them their large T-antigen and small t-antigen have been shown to possess oncogenic properties in cell culture and animal models, while similar functions are assumed for the large T- and small t-antigen of other human polyomaviruses. However, so far the Merkel cell polyomavirus seems to be the only human polyomavirus associated with cancer. The large T- and small t-antigen exert their tumorigenic effects through classical hallmarks of cancer: inhibiting tumor suppressors, activating tumor promoters, preventing apoptosis, inducing angiogenesis and stimulating metastasis. This review elaborates on the putative roles of human polyomaviruses in some of the emerging hallmarks of cancer. The reciprocal interactions between human polyomaviruses and the immune system response are discussed, a plausible role of polyomavirus-encoded and polyomavirus-induced microRNA in cancer is described, and the effect of polyomaviruses on energy homeostasis and exosomes is explored. Therapeutic strategies against these emerging hallmarks of cancer are also suggested.

  3. The Role of Merkel Cell Polyomavirus and Other Human Polyomaviruses in Emerging Hallmarks of Cancer

    PubMed Central

    Moens, Ugo; Rasheed, Kashif; Abdulsalam, Ibrahim; Sveinbjørnsson, Baldur

    2015-01-01

    Polyomaviruses are non-enveloped, dsDNA viruses that are common in mammals, including humans. All polyomaviruses encode the large T-antigen and small t-antigen proteins that share conserved functional domains, comprising binding motifs for the tumor suppressors pRb and p53, and for protein phosphatase 2A, respectively. At present, 13 different human polyomaviruses are known, and for some of them their large T-antigen and small t-antigen have been shown to possess oncogenic properties in cell culture and animal models, while similar functions are assumed for the large T- and small t-antigen of other human polyomaviruses. However, so far the Merkel cell polyomavirus seems to be the only human polyomavirus associated with cancer. The large T- and small t-antigen exert their tumorigenic effects through classical hallmarks of cancer: inhibiting tumor suppressors, activating tumor promoters, preventing apoptosis, inducing angiogenesis and stimulating metastasis. This review elaborates on the putative roles of human polyomaviruses in some of the emerging hallmarks of cancer. The reciprocal interactions between human polyomaviruses and the immune system response are discussed, a plausible role of polyomavirus-encoded and polyomavirus-induced microRNA in cancer is described, and the effect of polyomaviruses on energy homeostasis and exosomes is explored. Therapeutic strategies against these emerging hallmarks of cancer are also suggested. PMID:25866902

  4. [New, newer, newest human polyomaviruses: how far?].

    PubMed

    Us, Dürdal

    2013-04-01

    Polyomaviruses, classified in Polyomaviridae family, are non-enveloped small (40-45 nm) viruses with icosahedral symmetry and circular double-stranded DNA genome. Polyomaviruses can infect a variety of vertebrates including birds, rodents, cattle, monkeys and humans. The characteristics such as establishment of latent infections, reactivations during immunosuppression and oncogenic potencies render the human polyomaviruses (HPyVs) of considerable importance for public health. The first polyomavirus (Mouse polyomavirus) has been identified in 1953 as filterable tumor-causing agents in mice, followed by Simian vacuolating virus (SV40) isolated from rhesus monkey kidney cells that had been used for poliovirus vaccine preparation in 1960. Due to the known transforming capacity of SV40, it was initially thought that the incidence of cancer could increase following the administration of SV40-contaminated polio vaccines, however advanced studies yielded inconsistent results, without any evidence to conclude whether or not the contaminated polio vaccine caused cancer. Several studies have reported the detection of SV40 genome in some of the human tumors, as well as in the clinical samples of healthy subjects. In addition SV40 seropositivity was reported in human populations although in low rates (2-10%). These data have raised the possibility that SV40 infects humans and circulates in human populations unrelated to being exposed to the vaccine. The discovery of the first human polyomaviruses was in 1971 independently from each other, one was BK virus (BKPyV) isolated from the urine sample of a renal transplant patient, and the other was JC virus (JCPyV) isolated from the brain tissue of a patient with progressive multifocal leukoencephalopathy, and both were named after the patients' initials. BK and JC viruses were the only well-known human polyomaviruses throughout 36 years, however drammatical increase in number of newly identified human polyomaviruses was recorded in

  5. Merkel Cell Polyomavirus and Two Novel Polyomaviruses Are Chronically Shed from Human Skin

    PubMed Central

    Schowalter, Rachel M.; Pastrana, Diana V.; Pumphrey, Katherine A.; Moyer, Adam L.; Buck, Christopher B.

    2010-01-01

    Summary Mounting evidence supports the concept that Merkel cell polyomavirus (MCV) is a causal factor underlying most cases of a highly lethal form of skin cancer known as Merkel cell carcinoma. To explore the possibility that polyomaviruses commonly infect healthy human skin, we developed an improved rolling circle amplification (RCA) technique to isolate circular DNA viral genomes from skin swab specimens. Complete MCV genomes were recovered from 14/35 (40%) healthy adults, providing the first full-length, apparently wild-type cloned genomes for this polyomavirus species. RCA analysis also revealed the existence of two previously unknown polyomavirus species that we name human polyomavirus-6 (HPyV6) and HPyV7. Biochemical experiments show that polyomavirus DNA is shed from the skin in the form of assembled virions. A pilot serological study indicates that infection or co-infection with the three skin-tropic polyomaviruses is very common. Thus, at least three polyomavirus species are constituents of the human skin microbiome. PMID:20542254

  6. Age-specific seroprevalences of merkel cell polyomavirus, human polyomaviruses 6, 7, and 9, and trichodysplasia spinulosa-associated polyomavirus.

    PubMed

    Nicol, Jérôme T J; Robinot, Rémy; Carpentier, Audrey; Carandina, Giovanni; Mazzoni, Elisa; Tognon, Mauro; Touzé, Antoine; Coursaget, Pierre

    2013-03-01

    Six new human polyomaviruses have been identified since 2008 (Merkel cell polyomavirus [MCPyV], human polyomavirus 6 [HPyV6], HPyV7, HPyV9, trichodysplasia spinulosa polyomavirus [TSPyV], and Malawi polyomavirus [MWPyV]). The presence of specific antibodies against MCPyV, HPyV6, HPyV7, HPyV9, and TSPyV in 828 Italian subjects aged 1 to 100 years was investigated by virus-like particle-based enzyme-linked immunosorbent assays (ELISAs). The findings indicate that all of these new polyomaviruses circulate widely in humans, with seroprevalences in adulthood ranging from 39.4% for HPyV9 to 87.1% for MCPyV, and that primary exposure is most intense in childhood, with the exception of HPyV7 and HPyV9, for which the seroprevalence increased throughout life. The proportion of subjects with high antibody titers was found to increase with age for MCPyV and to decrease with age for TSPyV.

  7. A novel human polyomavirus closely related to the african green monkey-derived lymphotropic polyomavirus.

    PubMed

    Scuda, Nelly; Hofmann, Jörg; Calvignac-Spencer, Sébastien; Ruprecht, Klemens; Liman, Peter; Kühn, Joachim; Hengel, Hartmut; Ehlers, Bernhard

    2011-05-01

    We identified a novel human polyomavirus from a kidney transplant patient under immunosuppressive treatment, by use of a generic PCR. The genome of the virus was completely amplified and sequenced. In phylogenetic analyses, it appeared as the closest relative to the African green monkey-derived lymphotropic polyomavirus (LPV). Further investigation of clinical samples from immunocompromised patients with specific nested PCR revealed additional positive samples, indicating that the virus naturally infects humans. The virus was tentatively named human polyomavirus 9 (HPyV9). The previously observed seroreactivity to LPV in human populations might find a partial explanation in the circulation of HPyV9.

  8. Seroprevalence and cross-reactivity of human polyomavirus 9.

    PubMed

    Nicol, Jérôme T J; Touzé, Antoine; Robinot, Rémy; Arnold, Francoise; Mazzoni, Elisa; Tognon, Mauro; Coursaget, Pierre

    2012-08-01

    Many humans have antibodies against simian lymphotropic polyomavirus (LPyV), but its DNA has not been found in humans. Identification of human polyomavirus 9 (HPyV9) led us to compare the seroprevalence and cross-reactivity of LPyV and HpyV9. Results could indicate that humans who have antibodies against LPyV are infected by HPyV9.

  9. Early and late promoters of BK polyomavirus, Merkel cell polyomavirus, Trichodysplasia spinulosa-associated polyomavirus and human polyomavirus 12 are among the strongest of all known human polyomaviruses in 10 different cell lines.

    PubMed

    Moens, Ugo; Van Ghelue, Marijke; Ludvigsen, Maria; Korup-Schulz, Sarah; Ehlers, Bernhard

    2015-08-01

    Recently, 11 new human polyomaviruses (HPyVs) have been isolated and named KI, WU, Merkel cell polyomavirus (MCPyV), HPyV6,  -7,  -9,  -10 and  -12, Trichodysplasia spinulosa-associated polyomavirus (TSPyV), STLPyV and NJPyV-2013. Little is known about cell tropism of the novel HPyVs, and cell cultures allowing virus propagation are lacking. Because viral tropism partially depends on the interaction of cellular transcription factors with the viral promoter, we monitored the promoter activity of all known HPyVs. Therefore, we compared the relative early and late promoter activity of the BK polyomavirus (BKPyV) (WW strain) with the corresponding activities of the other HPyVs in 10 different cell lines derived from brain, colon, kidney, liver, lung, the oral cavity and skin. Our results show that the BKPyV, MCPyV, TSPyV and HPyV12 early promoters displayed the strongest activity in most cell lines tested, while the remaining HPyV had relative low early promoter activity. HPyV12 showed the highest late promoter activity of all HPyVs in most cell lines, but also the BKPyV, MCPyV and TSPyV late promoters belonged to the stronger ones among HPyVs. The HPyVs with weak early promoter activity had in general also weak late promoter activity, except for HPyV10 whose late promoter was relatively strong in six of the 10 cell lines. A 20 bp deletion in the promoter of an HPyV12 variant significantly affected both early and late promoter activity in most cell lines. In conclusion, our findings suggest which cell lines may be suitable for virus propagation and may give an indication of the cell tropism of the HPyVs.

  10. Human polyomavirus 9 infection in kidney transplant patients.

    PubMed

    van der Meijden, Els; Wunderink, Herman F; van der Blij-de Brouwer, Caroline S; Zaaijer, Hans L; Rotmans, Joris I; Bavinck, Jan Nico Bouwes; Feltkamp, Mariet C W

    2014-06-01

    Several human polyomaviruses of unknown prevalence and pathogenicity have been identified, including human polyomavirus 9 (HPyV9). To determine rates of HPyV9 infection among immunosuppressed patients, we screened serum samples from 101 kidney transplant patients in the Netherlands for HPyV9 DNA and seroreactivity. A total of 21 patients had positive results for HPyV9 DNA; positivity rates peaked at 3 months after transplantation, but the highest viral loads were measured just after transplantation. During 18 months of follow-up, HPyV9 seroprevalence increased from 33% to 46% among transplant patients; seroprevalence remained stable at ≈30% in a control group of healthy blood donors in whom no HPyV9 DNA was detected. Further analysis revealed an association between detection of HPyV9 and detection of BK polyomavirus but not of cytomegalovirus. Our data indicate that HPyV9 infection is frequent in kidney transplant patients, but the nature of infection-endogenous or donor-derived-and pathogenic potential of this virus remain unknown.

  11. Quantitation of Human Seroresponsiveness to Merkel Cell Polyomavirus

    PubMed Central

    Pastrana, Diana V.; Tolstov, Yanis L.; Becker, Jürgen C.; Moore, Patrick S.; Chang, Yuan; Buck, Christopher B.

    2009-01-01

    Merkel cell carcinoma (MCC) is a relatively uncommon but highly lethal form of skin cancer. A majority of MCC tumors carry DNA sequences derived from a newly identified virus called Merkel cell polyomavirus (MCV or MCPyV), a candidate etiologic agent underlying the development of MCC. To further investigate the role of MCV infection in the development of MCC, we developed a reporter vector-based neutralization assay to quantitate MCV-specific serum antibody responses in human subjects. Our results showed that 21 MCC patients whose tumors harbored MCV DNA all displayed vigorous MCV-specific antibody responses. Although 88% (42/48) of adult subjects without MCC were MCV seropositive, the geometric mean titer of the control group was 59-fold lower than the MCC patient group (p<0.0001). Only 4% (2/48) of control subjects displayed neutralizing titers greater than the mean titer of the MCV-positive MCC patient population. MCC tumors were found not to express detectable amounts of MCV VP1 capsid protein, suggesting that the strong humoral responses observed in MCC patients were primed by an unusually immunogenic MCV infection, and not by viral antigen expressed by the MCC tumor itself. The occurrence of highly immunogenic MCV infection in MCC patients is unlikely to reflect a failure to control polyomavirus infections in general, as seroreactivity to BK polyomavirus was similar among MCC patients and control subjects. The results support the concept that MCV infection is a causative factor in the development of most cases of MCC. Although MCC tumorigenesis can evidently proceed in the face of effective MCV-specific antibody responses, a small pilot animal immunization study revealed that a candidate vaccine based on MCV virus-like particles (VLPs) elicits antibody responses that robustly neutralize MCV reporter vectors in vitro. This suggests that a VLP-based vaccine could be effective for preventing the initial establishment of MCV infection. PMID:19750217

  12. The oncogenic potential of BK-polyomavirus is linked to viral integration into the human genome.

    PubMed

    Kenan, Daniel J; Mieczkowski, Piotr A; Burger-Calderon, Raquel; Singh, Harsharan K; Nickeleit, Volker

    2015-11-01

    It has been suggested that BK-polyomavirus is linked to oncogenesis via high expression levels of large T-antigen in some urothelial neoplasms arising following kidney transplantation. However, a causal association between BK-polyomavirus, large T-antigen expression and oncogenesis has never been demonstrated in humans. Here we describe an investigation using high-throughput sequencing of tumour DNA obtained from an urothelial carcinoma arising in a renal allograft. We show that a novel BK-polyomavirus strain, named CH-1, is integrated into exon 26 of the myosin-binding protein C1 gene (MYBPC1) on chromosome 12 in tumour cells but not in normal renal cells. Integration of the BK-polyomavirus results in a number of discrete alterations in viral gene expression, including: (a) disruption of VP1 protein expression and robust expression of large T-antigen; (b) preclusion of viral replication; and (c) deletions in the non-coding control region (NCCR), with presumed alterations in promoter feedback loops. Viral integration disrupts one MYBPC1 gene copy and likely alters its expression. Circular episomal BK-polyomavirus gene sequences are not found, and the renal allograft shows no productive polyomavirus infection or polyomavirus nephropathy. These findings support the hypothesis that integration of polyomaviruses is essential to tumourigenesis. It is likely that dysregulation of large T-antigen, with persistent over-expression in non-lytic cells, promotes cell growth, genetic instability and neoplastic transformation.

  13. Seroprevalence of human polyomavirus 9 and cross-reactivity to African green monkey-derived lymphotropic polyomavirus.

    PubMed

    Trusch, Franziska; Klein, Marcus; Finsterbusch, Tim; Kühn, Joachim; Hofmann, Jörg; Ehlers, Bernhard

    2012-04-01

    Human polyomavirus 9 (HPyV9) was discovered recently in immunocompromised patients and shown to be genetically closely related to B-lymphotropic polyomavirus (LPyV). No serological data are available for HPyV9, but human antibodies against LPyV have been reported previously. To investigate the seroepidemiology of HPyV9 and the sero-cross-reactivity between HPyV9 and LPyV, a capsomer-based IgG ELISA was established using the major capsid protein VP1 of HPyV9 and LPyV. VP1 of an avian polyomavirus was used as control. For HPyV9, a seroprevalence of 47 % was determined in healthy adults and adolescents (n = 328) and 20 % in a group of children (n =101). In both groups, the seroreactivities for LPyV were less frequent and the ELISA titres of LPyV were lower. Of the HPyV9-reactive sera, 47 % reacted also with LPyV, and the titres for both PyVs correlated. Sera from African green monkeys, the natural hosts of LPyV, reacted also with both HPyV9 and LPyV, but here the HPyV9 titres were lower. This potential sero-cross-reactivity between HPyV9 and LPyV was confirmed by competition assays, and it was hypothesized that the reactivity of human sera against LPyV may generally be due to cross-reactivity between HPyV9 and LPyV. The HPyV9 seroprevalence of liver transplant recipients and patients with neurological dysfunctions did not differ from that of age-matched controls, but a significantly higher seroprevalence was determined in renal and haematopoietic stem-cell transplant recipients, indicating that certain immunocompromised patient groups may be at a higher risk for primary infection with or for reactivation of HPyV9.

  14. BK Polyomavirus Genomic Integration and Large T Antigen Expression: Evolving Paradigms in Human Oncogenesis.

    PubMed

    Kenan, D J; Mieczkowski, P A; Latulippe, E; Côté, I; Singh, H K; Nickeleit, V

    2016-12-31

    Human polyomaviruses are ubiquitous, with primary infections that typically occur during childhood and subsequent latency that may last a lifetime. Polyomavirus-mediated disease has been described in immunocompromised patients; its relationship to oncogenesis is poorly understood. We present deep sequencing data from a high-grade BK virus-associated tumor expressing large T antigen. The carcinoma arose in a kidney allograft 6 years after transplantation. We identified a novel genotype 1a BK polyomavirus, called Chapel Hill BK polyomavirus 2 (CH-2), that was integrated into the BRE gene in chromosome 2 of tumor cells. At the chromosomal integration site, viral break points were found, disrupting late BK gene sequences encoding capsid proteins VP1 and VP2/3. Immunohistochemistry and in situ hybridization studies demonstrated that the integrated BK virus was replication incompetent. We propose that the BK virus CH-2 was integrated into the human genome as a concatemer, resulting in alterations of feedback loops and overexpression of large T antigen. Collectively, these findings support the emerging understanding that viral integration is a nearly ubiquitous feature in polyomavirus-associated malignancy and that unregulated large T antigen expression drives a proliferative state that is conducive to oncogenesis. Based on the current observations, we present an updated model of polyomavirus-mediated oncogenesis.

  15. Human polyomaviruses 6, 7, 9, 10 and Trichodysplasia spinulosa-associated polyomavirus in HIV-infected men.

    PubMed

    Wieland, Ulrike; Silling, Steffi; Hellmich, Martin; Potthoff, Anja; Pfister, Herbert; Kreuter, Alexander

    2014-04-01

    Recently, several novel human polyomaviruses (HPyVs) have been detected. HPyV6, 7, 9 and 10 are not associated with any disease so far. Trichodysplasia spinulosa (TS)-associated polyomavirus (TSPyV) can cause the rare skin disease TS. We have evaluated cutaneous DNA prevalence and viral loads of five HPyVs in HIV-infected men compared to healthy male controls. 449 forehead swabs were analysed by HPyV-specific real-time PCR. HPyV6, HPyV7, TSPyV and HPyV10 were found significantly more frequently on the skin of 210 HIV-infected compared to 239 HIV-negative men (HPyV6, 39.0 vs 27.6 %; HPyV7, 21.0 vs 13.4 %; TSPyV, 3.8 vs 0.8 %; HPyV10, 9.3 vs 3.4 %; P<0.05, respectively). HPyV9 was not detected. Multiple infections were more frequent in HIV-positive men, but HPyV-DNA loads did not differ significantly in both groups. In contrast to HPyV6, 7 and 10, TSPyV and HPyV9 do not seem to be a regular part of the human skin microbiome.

  16. Identification of a novel human polyomavirus in organs of the gastrointestinal tract.

    PubMed

    Korup, Sarah; Rietscher, Janita; Calvignac-Spencer, Sébastien; Trusch, Franziska; Hofmann, Jörg; Moens, Ugo; Sauer, Igor; Voigt, Sebastian; Schmuck, Rosa; Ehlers, Bernhard

    2013-01-01

    Polyomaviruses are small, non-enveloped viruses with a circular double-stranded DNA genome. Using a generic polyomavirus PCR targeting the VP1 major structural protein gene, a novel polyomavirus was initially identified in resected human liver tissue and provisionally named Human Polyomavirus 12 (HPyV12). Its 5033 bp genome is predicted to encode large and small T antigens and the 3 structural proteins VP1, VP2 and VP3. Phylogenetic analyses did not reveal a close relationship to any known human or animal polyomavirus. Investigation of organs, body fluids and excretions of diseased individuals and healthy subjects with both HPyV12-specific nested PCR and quantitative real-time PCR revealed additional virus-positive samples of resected liver, cecum and rectum tissues and a positive fecal sample. A capsomer-based IgG ELISA was established using the major capsid protein VP1 of HPyV12. Seroprevalences of 23% and 17%, respectively, were determined in sera from healthy adults and adolescents and a pediatric group of children. These data indicate that the virus naturally infects humans and that primary infection may already occur in childhood.

  17. Seroepidemiology of Human Polyomaviruses in a US Population

    PubMed Central

    Gossai, Anala; Waterboer, Tim; Nelson, Heather H.; Michel, Angelika; Willhauck-Fleckenstein, Martina; Farzan, Shohreh F.; Hoen, Anne G.; Christensen, Brock C.; Kelsey, Karl T.; Marsit, Carmen J.; Pawlita, Michael; Karagas, Margaret R.

    2016-01-01

    Polyomaviruses (PyV) are potentially tumorigenic in humans. However, limited data exist on the population seroprevalence of PyVs and individual characteristics that relate to seropositivity. Using multiplex serology, we determined the seroprevalence of 10 human PyVs (BK, JC, KI, WU, MCV, HPyV6, HPyV7, TSV, HPyV9, and HPyV10) among controls from a population-based skin cancer case-control study (n = 460) conducted in New Hampshire between 1993 and 1995. On a subset of participants (n = 194), methylation at CpG dinucleotides across the genome was measured in peripheral blood using the Illumina Infinium HumanMethylation27 BeadChip array (Illumina Inc., San Diego, California), from which lymphocyte subtype proportions were inferred. All participants were seropositive for at least 1 PyV, with seroprevalences ranging from 17.6% (HPyV9) to 99.1% (HPyV10). Seropositivity to JC, MCV, and HPyV7 increased with age. JC and TSV seropositivity were more common among men than among women. Smokers were more likely to be HPyV9-seropositive but MCV-seronegative, and HPyV7 seropositivity was associated with prolonged glucocorticoid use. Based on DNA methylation profiles, differences were observed in CD8-positive T- and B-cell proportions by BK, JC, and HPyV9 seropositivity. Our findings suggest that PyV seropositivity is common in the United States and varies by sociodemographic and biological characteristics, including those related to immune function. PMID:26667254

  18. Malawi polyomavirus is a prevalent human virus that interacts with known tumor suppressors.

    PubMed

    Berrios, Christian; Jung, Joonil; Primi, Blake; Wang, Michael; Pedamallu, Chandrasekhar; Duke, Fujiko; Marcelus, Christina; Cheng, Jingwei; Garcea, Robert L; Meyerson, Matthew; DeCaprio, James A

    2015-01-01

    Malawi polyomavirus (MWPyV) is a recently identified human polyomavirus. Serology for MWPyV VP1 indicates that infection frequently occurs in childhood and reaches a prevalence of 75% in adults. The MWPyV small T antigen (ST) binds protein phosphatase 2A (PP2A), and the large T antigen (LT) binds pRb, p107, p130, and p53. However, the MWPyV LT was less stable than the simian virus 40 (SV40) LT and was unable to promote the growth of normal cells. This report confirms that MWPyV is a widespread human virus expressing T antigens with low transforming potential.

  19. High diversity of human polyomaviruses in environmental and clinical samples in Argentina: Detection of JC, BK, Merkel-cell, Malawi, and human 6 and 7 polyomaviruses.

    PubMed

    Torres, Carolina; Barrios, Melina Elizabeth; Cammarata, Robertina Viviana; Cisterna, Daniel Marcelo; Estrada, Tatiana; Martini Novas, Sergio; Cahn, Pedro; Blanco Fernández, María Dolores; Mbayed, Viviana Andrea

    2016-01-15

    New human polyomaviruses have been recently described. The aim of this work was to detect and characterize human polyomaviruses circulating in Argentina by recovering viruses from environmental and sewage samples and evaluating their potential role as viral indicators of human waste contamination. Analysis was performed in a wider context including viruses from clinical samples from an immunocompromised population. River water and sewage samples were analyzed as a strategy to study the molecular epidemiology of viruses excreted by millions of people. Samples belonged to the Matanza-Riachuelo River (2005-2006: n=25 and 2012: n=20) and sewage from Buenos Aires city and suburbs (2011 and 2013: n=24). Viral detection was performed by PCR and the amplified viral genomes were characterized by phylogenetic analysis. Polyomaviruses were detected in 95.8% of sewage samples, identifying BKPyV (87.5%), JCPyV (83.3%), MCPyV (8.3%) and HPyV6 (8.3%). Besides, one sample collected in 2009 resulted positive for HPyV7. In 2005-2006, polyomaviruses were detected in 84.0% of river water samples, with the highest detection for MCPyV (52.0%), followed by BKPyV (44.0%), JCPyV (20.0%) and MWPyV (4.0%). In 2012, polyomaviruses were detected in 85.0% of river samples, finding JCPyV (85.0%), BKPyV (75.0%), MCPyV (25.0%) and HPyV6 (25.0%). Also, polyomaviruses, including JCPyV, BKPyV and MCPyV, were detected in 63.2% of urine samples from patients infected with HIV (n=19). Characterization indicated the coexistence of different genotypes and variants for each virus, particularly in sewage. MCPyV sequences (the only sequences from Argentina) formed a monophyletic group with the single sequence available for South America (French Guiana). The high level of detection and viral diversity found by environmental surveillance, which involved the characterization of viruses not previously described in South America, reinforces the usefulness of this approach to monitor viral contamination and

  20. Polyomavirus-associated nephritis in 2 horses.

    PubMed

    Jennings, S H; Wise, A G; Nickeleit, V; Maes, R K; Cianciolo, R E; Del Piero, F; Law, J M; Kim, Y; McCalla, A C; Breuhaus, B A; Roberts, M C; Linder, K E

    2013-09-01

    Polyomaviruses produce latent and asymptomatic infections in many species, but productive and lytic infections are rare. In immunocompromised humans, polyomaviruses can cause tubulointerstitial nephritis, demyelination, or meningoencephalitis in the central nervous system and interstitial pneumonia. This report describes 2 Standardbred horses with tubular necrosis and tubulointerstitial nephritis associated with productive equine polyomavirus infection that resembles BK polyomavirus nephropathy in immunocompromised humans.

  1. Role of the Human Polyomavirus, BKV, in Prostate Cancer

    DTIC Science & Technology

    2004-08-01

    polyomavirus and late regions, and encodes five major proteins nephropathy (Arthur et al., 1986; Hiraoka et al., 1991; (Figure 1; Seif et al., 1979...specimens was first ascertained by PCR (45 cycles) with the f-globin primers. stained with hematoxylin and cosin to confirm diagnosis . The The... pregnancy , or HIV infection, high throughout the rest of the time course. levels of BKV replication are observed in the kidneys and The next step in the BKV

  2. Role of Human Polyomavirus Bkv in Prostate Cancer

    DTIC Science & Technology

    2007-12-01

    been surgically removed due to prostate cancer diagnosis . A normal prostate is defined as prostate that has been removed either during autopsy or by...immunosuppressed transplant patients, in whom it is associated with haemorrhagic cystitis and polyomavirus nephropathy (5, 35, 58, 70). BKV transforms rodent cells...cystoprostatectomy specimens from bladder cancer patients with the diagnosis of muscle invasive high grade urothelial carcinoma, with no prostate cancer histology

  3. Human bocavirus and KI/WU polyomaviruses in pediatric intensive care patients.

    PubMed

    van de Pol, Alma C; Wolfs, Tom F W; Jansen, Nicolaas J G; Kimpen, Jan L L; van Loon, Anton M; Rossen, John W A

    2009-03-01

    We evaluated the prevalence of human bocavirus and KI and WU polyomaviruses in pediatric intensive care patients with and without lower respiratory tract infection (LRTI). The prevalence of these viruses was 5.1%, 0%, and 2.6%, respectively, in children with LRTI and 4.8%, 4.8%, and 2.4%, respectively, in those without LRTI.

  4. Detection of polyomavirus major capsid antigen (VP-1) in human pilomatricomas.

    PubMed

    Sanjuán, Norberto A; Símula, Silvina; Casas, José; Woscoff, Alberto

    2010-01-01

    The family Polyomaviridae is composed of small, non-enveloped, double-stranded DNA viruses widely used to study cell transformation in vitro and tumor induction in vivo. The development of pilomatricomas in mice experimentally infected with polyomavirus led us to detect the viral major capsid protein VP-1 in human pilomatricomas. This tumor, even uncommon, is one of the most frequent benign hair follicle tumors in humans and is composed of proliferating matrix cells that undergo keratinization, and form cystic neoplasms. The detection of VP-1 was performed using the peroxidase-antiperoxidase technique in paraffin-embedded slides with a specific primary serum. Adjacent slides treated with normal rabbit serum as a primary were employed as internal control. Positive and negative controls were also employed as well as slides of lesions caused by human papillomavirus to rule out any unspecific cross-reactivity. In 4 out of 10 cases polyomavirus VP-1 was clearly detected in nuclei of human pilomatricomas proliferating cells, in a patchy pattern of distribution. The controls confirmed the specificity of the immunocytochemical procedure. These results could indicate either an eventual infection of the virus in already developed tumors or alternatively, a direct involvement of polyomavirus in the pathogenesis of some pilomatricomas. The recent discovery of a new human polyomavirus associated with Merkel cell carcinomas has been a strong contribution to better understand the pathogenesis of some human uncommon skin cancers. Hopefully the results reported in this work will encourage further research on the role of polyomavirus in other human skin neoplasms.

  5. A prospective study of human polyomavirus infection in pregnancy.

    PubMed

    Coleman, D V; Wolfendale, M R; Daniel, R A; Dhanjal, N K; Gardner, S D; Gibson, P E; Field, A M

    1980-07-01

    Urine samples from 1,235 pregnant women were examined by light microscopy for cytologic evidence of virus infection. Smears of urine sediment from 40 women (3.2%) were observed to contain inclusion-bearing cells; polyomavirus infection was confirmed by virologic methods in 24 (60%). A polyomavirus was isolated from 12 women. Five isolates were identified as JC virus and one as BK virus. Another isolate designated AS virus appeared to be unique. Serologic studies on the 40 women were consistent with a high frequency of reactivation of JC virus, and virus excretion was related to gestation. The evidence suggests that selective excretion of JC virus may occur in pregnancy. Among 390 pregnant women without inclusion-bearing cells in their urine, 78 (20%) had a high or rising titer of serum antibody to JC or BK virus or both, a result suggesting virus reactivation, but virus excretion was not detected. In contrast to other reports, no evidence was found for transmission of BK virus to the fetus.

  6. In Vitro and In Vivo Models for the Study of Human Polyomavirus Infection

    PubMed Central

    Barth, Heidi; Solis, Morgane; Kack-Kack, Wallys; Soulier, Eric; Velay, Aurélie; Fafi-Kremer, Samira

    2016-01-01

    Developments of genome amplification techniques have rapidly expanded the family of human polyomaviruses (PyV). Following infection early in life, PyV persist in their hosts and are generally of no clinical consequence. High-level replication of PyV can occur in patients under immunosuppressive or immunomodulatory therapy and causes severe clinical entities, such as progressive multifocal leukoencephalopathy, polyomavirus-associated nephropathy or Merkel cell carcinoma. The characterization of known and newly-discovered human PyV, their relationship to human health, and the mechanisms underlying pathogenesis remain to be elucidated. Here, we summarize the most widely-used in vitro and in vivo models to study the PyV-host interaction, pathogenesis and anti-viral drug screening. We discuss the strengths and limitations of the different models and the lessons learned. PMID:27782080

  7. First Detection of Human Papillomaviruses and Human Polyomaviruses in River Waters in Italy.

    PubMed

    Iaconelli, M; Petricca, S; Libera, S Della; Di Bonito, P; La Rosa, G

    2015-12-01

    Waterborne exposure to human viruses is possible through contact with contaminated water environments and can result in infections associated with a wide range of illnesses, including gastrointestinal, respiratory, ear, ocular, and skin infections. Recently, the occurrence in water environments of two groups of human viruses-both known with oncogenic potential, human polyomaviruses (HPyVs) and papillomaviruses (HPVs)-has been reported worldwide. These viruses, responsible for highly prevalent infections worldwide, have recently been proposed as potentially emerging waterborne pathogens. The objective of the present study was to examine the occurrence of HPyVs and HPVs in surface waters, by monitoring two rivers in Northwestern Italy, by nested PCR assays and sequencing. HPyVs (JC, BK, and Merkel cell polyomavirus) were detected in 10/25 (40%) samples. HPVs (HPV8, 17, 21, 25, 32, 80, 99, 105, and putative new HPVs) were identified in 14/25 (56%) river samples. The number of HPV DNA copies in waters was measured by quantitative real-time PCR. To our knowledge, this is the first detection and quantification of HPVs in surface waters. The possibility that HPyVs and HPVs can be transmitted by the waterborne route deserves to be explored in future studies.

  8. The human polyomaviruses KI and WU: virological background and clinical implications.

    PubMed

    Babakir-Mina, Muhammed; Ciccozzi, Massimo; Perno, Carlo Federico; Ciotti, Marco

    2013-08-01

    In 2007, two novel polyomaviruses KI and WU were uncovered in the respiratory secretions of children with acute respiratory symptoms. Seroepidemiological studies showed that infection by these viruses is widespread in the human population. Following these findings, different biological specimens and body compartments have been screened by real-time PCR in the attempt to establish a pathogenetic role for KI polyomavirus (KIPyV) and WU polyomavirus (WUPyV) in human diseases. Although both viruses have been found mainly in respiratory tract samples of immunocompromised patients, a clear causative link with the respiratory disease has not been established. Indeed, the lack of specific clinical or radiological findings, the frequent co-detection with other respiratory pathogens, the detection in subjects without signs or symptoms of respiratory disease, and the variability of the viral loads measured did not allow drawing a definitive conclusion. Prospective studies carried out on a large sample size including both immunocompromised and immunocompetent patients with and without respiratory symptoms are needed. Standardized quantitative real-time PCR methods, definition of a clear clinical cutoff value, timing in the collection of respiratory samples, are also crucial to understand the pathogenic role, if any, of KIPyV and WUPyV in human pathology.

  9. Merkel Cell Polyomavirus: Molecular Insights into the Most Recently Discovered Human Tumour Virus

    PubMed Central

    Stakaitytė, Gabrielė; Wood, Jennifer J.; Knight, Laura M.; Abdul-Sada, Hussein; Adzahar, Noor Suhana; Nwogu, Nnenna; Macdonald, Andrew; Whitehouse, Adrian

    2014-01-01

    A fifth of worldwide cancer cases have an infectious origin, with viral infection being the foremost. One such cancer is Merkel cell carcinoma (MCC), a rare but aggressive skin malignancy. In 2008, Merkel cell polyomavirus (MCPyV) was discovered as the causative agent of MCC. It is found clonally integrated into the majority of MCC tumours, which require MCPyV oncoproteins to survive. Since its discovery, research has begun to reveal the molecular virology of MCPyV, as well as how it induces tumourigenesis. It is thought to be a common skin commensal, found at low levels in healthy individuals. Upon loss of immunosurveillance, MCPyV reactivates, and a heavy viral load is associated with MCC pathogenesis. Although MCPyV is in many ways similar to classical oncogenic polyomaviruses, such as SV40, subtle differences are beginning to emerge. These unique features highlight the singular position MCPyV has as the only human oncogenic polyomavirus, and open up new avenues for therapies against MCC. PMID:24978434

  10. Genome Sequence of a Central Chimpanzee-Associated Polyomavirus Related to BK and JC Polyomaviruses, Pan troglodytes troglodytes Polyomavirus 1.

    PubMed

    Madinda, Nadège F; Robbins, Martha M; Boesch, Christophe; Leendertz, Fabian H; Ehlers, Bernhard; Calvignac-Spencer, Sébastien

    2015-09-03

    We amplified and sequenced the genome of a polyomavirus infecting a central chimpanzee (Pan troglodytes troglodytes). This virus, which is closely related to BK and JC polyomaviruses, may help shed a new light on these human pathogens' evolutionary history.

  11. Multiple Skin Cancers in a Renal Transplant Recipient: A Patient Report with Analyses of Human Papillomavirus and Human Polyomavirus Infection

    PubMed Central

    Kaneda, Tokinobu; Matsushita, Michiko; Iwasaki, Takeshi; Ishiguro, Naoko; Koide, Takashi; Hayashi, Kazuhiko; Kitamura, Yukisato

    2015-01-01

    Skin cancer is an important complication in renal transplant recipients. Associations of transplant-related skin tumor with ultraviolet radiation, age at transplantation, type of immunosuppressant drug administered, and viral infection have been reported; however, the details remain unclear. We report a 61-year-old man who had underwent renal transplantation at 38 years of age and developed multiple skin tumors or squamous cell carcinomas (SCCs). Polymerase chain reaction (PCR) analyses of the patient’s 12 tumors for viral DNAs of cutaneous or mucosal human papillomavirus (HPV) and 6 human polyomaviruses (MCPyV, trichodysplasia spinulosa-associated, BK, JC, KI and WU polyomaviruses) only detected cutaneous HPV-DNA in only 5 of the tumors; no other viruses were detected. Real-time PCR showed high loads of cutaneous HPV in 3 SCCs and very low loads of MCPyV in 9. Immunohistochemistry revealed no tumor cell expression for MCPyV-large T-antigen or mucosal HPV. Our report not only reconfirmed the association of cutaneous HPV5 with skin cancer in renal transplant recipients in previous studies but also showed no relevant association of 6 human polyomaviruses and mucosal HPV with skin tumors. PMID:26538801

  12. Pseudovirus mimics cell entry and trafficking of the human polyomavirus JCPyV

    PubMed Central

    Gee, Gretchen V.; O’Hara, Bethany A.; Derdowski, Aaron; Atwood, Walter J.

    2013-01-01

    The normally asymptomatic human polyomavirus, JCPyV, is the causative agent of a rare but fatal demyelinating disease known as Progressive Multifocal Leukoencephalopathy (PML). Individuals at risk for developing PML include those with AIDS, with other underlying immunosuppressive diseases, and in patients treated with immunomodulatory regimens. Drugs to prevent viral reactivation in the setting of immunosuppression or immunomodulation could be used to sustain lives. Development of such drugs has been impeded by the difficulty of growing and studying the virus. We sought to develop a more efficient method for screening drugs that inhibit viral infection. Pseudovirus models have been developed which may be of use in pharmaceutical research. The use of pseudoviruses as models for viral infection is dependent on them using similar pathways for infection as virus. We screened known inhibitors of viral entry for their ability to block pseudovirus infection. Here we show that the pseudovirus based on the human polyomavirus JCPyV recapitulates virus binding, entry and trafficking. This system can be used for high-throughput screening of antiviral drugs. PMID:24100235

  13. Pseudovirus mimics cell entry and trafficking of the human polyomavirus JCPyV.

    PubMed

    Gee, Gretchen V; O'Hara, Bethany A; Derdowski, Aaron; Atwood, Walter J

    2013-12-26

    The normally asymptomatic human polyomavirus, JCPyV, is the causative agent of a rare but fatal demyelinating disease known as progressive multifocal leukoencephalopathy (PML). Individuals at risk for developing PML include those with AIDS, with other underlying immunosuppressive diseases, and in patients treated with immunomodulatory regimens. Drugs to prevent viral reactivation in the setting of immunosuppression or immunomodulation could be used to sustain lives. Development of such drugs has been impeded by the difficulty of growing and studying the virus. We sought to develop a more efficient method for screening drugs that inhibit viral infection. Pseudovirus models have been developed which may be of use in pharmaceutical research. The use of pseudoviruses as models for viral infection is dependent on them using similar pathways for infection as virus. We screened known inhibitors of viral entry for their ability to block pseudovirus infection. Here we show that the pseudovirus based on the human polyomavirus JCPyV recapitulates virus binding, entry and trafficking. This system can be used for high-throughput screening of antiviral drugs.

  14. Human Polyomavirus Receptor Distribution in Brain Parenchyma Contrasts with Receptor Distribution in Kidney and Choroid Plexus

    PubMed Central

    Haley, Sheila A.; O'Hara, Bethany A.; Nelson, Christian D.S.; Brittingham, Frances L.P.; Henriksen, Kammi J.; Stopa, Edward G.; Atwood, Walter J.

    2016-01-01

    The human polyomavirus, JCPyV, is the causative agent of progressive multifocal leukoencephalopathy, a rare demyelinating disease that occurs in the setting of prolonged immunosuppression. After initial asymptomatic infection, the virus establishes lifelong persistence in the kidney and possibly other extraneural sites. In rare instances, the virus traffics to the central nervous system, where oligodendrocytes, astrocytes, and glial precursors are susceptible to lytic infection, resulting in progressive multifocal leukoencephalopathy. The mechanisms by which the virus traffics to the central nervous system from peripheral sites remain unknown. Lactoseries tetrasaccharide c (LSTc), a pentasaccharide containing a terminal α2,6–linked sialic acid, is the major attachment receptor for polyomavirus. In addition to LSTc, type 2 serotonin receptors are required for facilitating virus entry into susceptible cells. We studied the distribution of virus receptors in kidney and brain using lectins, antibodies, and labeled virus. The distribution of LSTc, serotonin receptors, and virus binding sites overlapped in kidney and in the choroid plexus. In brain parenchyma, serotonin receptors were expressed on oligodendrocytes and astrocytes, but these cells were negative for LSTc and did not bind virus. LSTc was instead found on microglia and vascular endothelium, to which virus bound abundantly. Receptor distribution was not changed in the brains of patients with progressive multifocal leukoencephalopathy. Virus infection of oligodendrocytes and astrocytes during disease progression is LSTc independent. PMID:26056932

  15. Human polyomavirus receptor distribution in brain parenchyma contrasts with receptor distribution in kidney and choroid plexus.

    PubMed

    Haley, Sheila A; O'Hara, Bethany A; Nelson, Christian D S; Brittingham, Frances L P; Henriksen, Kammi J; Stopa, Edward G; Atwood, Walter J

    2015-08-01

    The human polyomavirus, JCPyV, is the causative agent of progressive multifocal leukoencephalopathy, a rare demyelinating disease that occurs in the setting of prolonged immunosuppression. After initial asymptomatic infection, the virus establishes lifelong persistence in the kidney and possibly other extraneural sites. In rare instances, the virus traffics to the central nervous system, where oligodendrocytes, astrocytes, and glial precursors are susceptible to lytic infection, resulting in progressive multifocal leukoencephalopathy. The mechanisms by which the virus traffics to the central nervous system from peripheral sites remain unknown. Lactoseries tetrasaccharide c (LSTc), a pentasaccharide containing a terminal α2,6-linked sialic acid, is the major attachment receptor for polyomavirus. In addition to LSTc, type 2 serotonin receptors are required for facilitating virus entry into susceptible cells. We studied the distribution of virus receptors in kidney and brain using lectins, antibodies, and labeled virus. The distribution of LSTc, serotonin receptors, and virus binding sites overlapped in kidney and in the choroid plexus. In brain parenchyma, serotonin receptors were expressed on oligodendrocytes and astrocytes, but these cells were negative for LSTc and did not bind virus. LSTc was instead found on microglia and vascular endothelium, to which virus bound abundantly. Receptor distribution was not changed in the brains of patients with progressive multifocal leukoencephalopathy. Virus infection of oligodendrocytes and astrocytes during disease progression is LSTc independent.

  16. Serological cross-reactivity between Merkel cell polyomavirus and two closely related chimpanzee polyomaviruses.

    PubMed

    Nicol, Jérôme T J; Liais, Etienne; Potier, Romain; Mazzoni, Elisa; Tognon, Mauro; Coursaget, Pierre; Touzé, Antoine

    2014-01-01

    Phylogenetic analyses based on the major capsid protein sequence indicate that Merkel cell polyomavirus (MCPyV) and chimpanzee polyomaviruses (PtvPyV1, PtvPyV2), and similarly Trichodysplasia spinulosa-associated polyomavirus (TSPyV) and the orangutan polyomavirus (OraPyV1) are closely related. The existence of cross-reactivity between these polyomaviruses was therefore investigated. The findings indicated serological identity between the two chimpanzee polyomaviruses investigated and a high level of cross-reactivity with Merkel cell polyomavirus. In contrast, cross-reactivity was not observed between TSPyV and OraPyV1. Furthermore, specific antibodies to chimpanzee polyomaviruses were detected in chimpanzee sera by pre-incubation of sera with the different antigens, but not in human sera.

  17. Human polyomavirus 7-associated pruritic rash and viremia in transplant recipients.

    PubMed

    Ho, Jonhan; Jedrych, Jaroslaw J; Feng, Huichen; Natalie, August A; Grandinetti, Lisa; Mirvish, Ezra; Crespo, Maria M; Yadav, Dhiraj; Fasanella, Kenneth E; Proksell, Siobhan; Kuan, Shih-Fan; Pastrana, Diana V; Buck, Christopher B; Shuda, Yoko; Moore, Patrick S; Chang, Yuan

    2015-05-15

    Human polyomavirus 7 (HPyV7) is one of 11 HPyVs recently discovered through genomic sequencing technologies. Two lung transplant recipients receiving immunosuppressive therapy developed pruritic, brown plaques on the trunk and extremities showing a distinctive epidermal hyperplasia with virus-laden keratinocytes containing densely packed 36-45-nm icosahedral capsids. Rolling circle amplification and gradient centrifugation testing were positive for encapsidated HPyV7 DNA in skin and peripheral blood specimens from both patients, and HPyV7 early and capsid proteins were abundantly expressed in affected tissues. We describe for the first time that HPyV7 is associated with novel pathogenicity in some immunosuppressed individuals.

  18. The Natural History of Human Polyomaviruses and Herpesviruses in Early Life--The Rhea Birth Cohort in Greece.

    PubMed

    Karachaliou, Marianna; Waterboer, Tim; Casabonne, Delphine; Chalkiadaki, Georgia; Roumeliotaki, Theano; Michel, Angelika; Stiakaki, Eftichia; Chatzi, Leda; Pawlita, Michael; Kogevinas, Manolis; de Sanjose, Silvia

    2016-04-01

    Sparse data exist on the patterns and determinants of acquisition of polyomaviruses and herpesviruses in childhood. We measured immunoglobulin G seroreactivity against 10 polyomaviruses (BKPyV, JCPyV, KIPyV, WUPyV, MCPyV, HPyV6, HPyV7, TSPyV, HPyV9, HPyV10) and 5 herpesviruses (Epstein Barr virus (EBV), cytomegalovirus (CMV), herpes simplex virus types 1 and 2, human herpesvirus 8) using multiplex serology on blood samples collected at birth (cord blood, n = 626) and at follow-up at 3 years (n = 81) and 4 years (n = 690) of age among the Rhea birth cohort recruited in Greece from pregnant women in 2007-2008. We used Poisson regression with robust variance to identify determinants of seropositivity at age 4. Seroprevalence of polyomaviruses ranged from 38.5% to 99.8% in cord blood and from 20.9% to 82.3% at age 4. Seroprevalence of EBV, CMV, herpes simplex virus types 1 and 2, and human herpesvirus 8 was 99.4%, 74.9%, 26.2%, 8.0%, and 1.6% in cord blood and 52.5%, 25.8%, 3.6%, 1.4%, and 0% at age 4, respectively. Determinants of seropositivity at age 4 were cord seropositivity (JCPyV, HPyV7, HPyV10, CMV), vaginal delivery (HPyV10), breastfeeding (CMV), younger age at day-care entry (BKPyV, KIPyV, WUPyV, TSPyV, HPyV10, HPyV9, EBV, CMV), and swimming pool attendance (BKPyV, KIPyV, WUPyV, HPyV10). Television viewing, parental stress, and hygiene practices were inversely associated with the seroprevalence of polyomaviruses and herpesviruses.

  19. Complete genomic sequence of a new Human polyomavirus 9 strain with an altered noncoding control region.

    PubMed

    Lednicky, John A; Butel, Janet S; Luetke, Maya C; Loeb, Julia C

    2014-12-01

    A complete Human polyomavirus 9 (HPyV9) genome, designated HPyV9 UF-1, was amplified by rolling circle DNA amplification from DNA extracted from the peripheral blood mononuclear cells (PBMC) of an AIDS patient. The noncoding control (enhancer/promoter) region (NCCR) of HPyV9 UF-1 has one less AML-1a binding site and three more potential Sp1/GC box binding sites than the NCCRs of two previously described HPyV9 genomes. Nucleotide polymorphisms within the coding regions result in two amino acid differences in the deduced VP2 and VP3 proteins of HPyV9 UF-1 relative to those of the two previously described HPyV9 genomes. Exhaustive attempts to detect HPyV9 in DNA samples extracted from the PBMC of 40 healthy humans and 9 other AIDS patients were unsuccessful, highlighting the need for improved search strategies and optimal specimens for the detection of HPyV9 in humans.

  20. Longitudinal study of seroprevalence and serostability of the human polyomaviruses JCV and BKV in organ transplant recipients.

    PubMed

    Antonsson, Annika; Pawlita, Michael; Feltkamp, Mariet C W; Bouwes Bavinck, Jan N; Euvrard, Sylvie; Harwood, Catherine A; Naldi, Luigi; Nindl, Ingo; Proby, Charlotte M; Neale, Rachel E; Waterboer, Tim

    2013-02-01

    The human polyomaviruses BKV and JCV cause mostly subclinical infections in childhood. Systemical immunosuppression after organ transplantation can lead to reactivation of persistent polyomavirus infections which may cause rejection of the transplanted organ. BKV and JCV seroprevalence and serostability was measured in 441 European solid organ transplanted recipients. Baseline samples were collected on average 24 days post-transplantation and sera were then collected over an 18 months follow-up period on up to six different time points. The overall seroprevalence at baseline for BKV was 97% with very little change over time. Prevalence for JCV was 76% at baseline and increased to 80% at the end of follow-up. BKV seroprevalence was highest in the youngest age group (100%) and decreased with increasing age (92% in the oldest age group; P < 0.0001), while JCV increased with age (69% vs. 81%; P = 0.020). Antibody reactivities for both BKV and JCV increased significantly with time (P = 0.0002 and P < 0.0001, respectively). Among the 406 patients with several samples, 94% were stably seropositive for BKV and 1% remained seronegative during the follow-up. JCV antibody stability was somewhat lower: 67% remained stably seropositive and 13% seronegative. While seroprevalence of BKV and JCV decrease and increase with age, respectively, both polyomaviruses showed significant increasing antibody reactivity over time in organ transplanted recipients at the onset of immunosuppression.

  1. The human alpha defensin HD5 neutralizes JC polyomavirus infection by reducing endoplasmic reticulum traffic and stabilizing the viral capsid.

    PubMed

    Zins, Stephen R; Nelson, Christian D S; Maginnis, Melissa S; Banerjee, Rahul; O'Hara, Bethany A; Atwood, Walter J

    2014-01-01

    Progressive multifocal leukoencephalopathy (PML) is a fatal disease with limited treatment options, both clinically and in the research pipeline. Potential therapies would target and neutralize its etiologic agent, JC polyomavirus (JCPyV). The innate immune response to JCPyV infection has not been studied, and little is known about the initial host response to polyomavirus infection. This study examined the ability of a human alpha defensin, HD5, to neutralize JCPyV infection in human fetal glial cells. We show that HD5, by binding to the virion, blocks infection. The JCPyV-HD5 complexes bind to and enter host cells but are reduced in their ability to reach the endoplasmic reticulum (ER), where virions are normally uncoated. Furthermore, HD5 binding to the virion stabilizes the capsid and prevents genome release. Our results show that HD5 neutralizes JCPyV infection at an early postentry step in the viral life cycle by stabilizing the viral capsid and disrupting JCPyV trafficking. This study provides a naturally occurring platform for developing antivirals to treat PML and also expands on the known capabilities of human defensins.

  2. The Human Alpha Defensin HD5 Neutralizes JC Polyomavirus Infection by Reducing Endoplasmic Reticulum Traffic and Stabilizing the Viral Capsid

    PubMed Central

    Zins, Stephen R.; Nelson, Christian D. S.; Maginnis, Melissa S.; Banerjee, Rahul; O'Hara, Bethany A.

    2014-01-01

    Progressive multifocal leukoencephalopathy (PML) is a fatal disease with limited treatment options, both clinically and in the research pipeline. Potential therapies would target and neutralize its etiologic agent, JC polyomavirus (JCPyV). The innate immune response to JCPyV infection has not been studied, and little is known about the initial host response to polyomavirus infection. This study examined the ability of a human alpha defensin, HD5, to neutralize JCPyV infection in human fetal glial cells. We show that HD5, by binding to the virion, blocks infection. The JCPyV-HD5 complexes bind to and enter host cells but are reduced in their ability to reach the endoplasmic reticulum (ER), where virions are normally uncoated. Furthermore, HD5 binding to the virion stabilizes the capsid and prevents genome release. Our results show that HD5 neutralizes JCPyV infection at an early postentry step in the viral life cycle by stabilizing the viral capsid and disrupting JCPyV trafficking. This study provides a naturally occurring platform for developing antivirals to treat PML and also expands on the known capabilities of human defensins. PMID:24198413

  3. Human Genome Diversity workshop 1

    SciTech Connect

    1992-12-31

    The Human Genome Diversity Project (HGD) is an international interdisciplinary program whose goal is to reveal as much as possible about the current state of genetic diversity among humans and the processes that were responsible for that diversity. Classical premolecular techniques have already proved that a significant component of human genetic variability lies within populations rather than among them. New molecular techniques will permit a dramatic increase in the resolving power of genetic analysis at the population level. Recent social changes in many parts of the world threaten the identity of a number of populations that may be extremely important for understanding human evolutionary history. It is therefore urgent to conduct research on human variation in these areas, while there is still time. The plan is to identify the most representative descendants of ancestral human populations worldwide and then to preserve genetic records of these populations. This is a report of the Population Genetics Workshop (Workshop 1), the first of three to be held to plan HGD, which was focused on sampling strategies and analytic methods from population genetics. The topics discussed were sampling and population structure; analysis of populations; drift versus natural selection; modeling migration and population subdivision; and population structure and subdivision.

  4. JC Polyomavirus Infection Is Strongly Controlled by Human Leucocyte Antigen Class II Variants

    PubMed Central

    Sundqvist, Emilie; Buck, Dorothea; Warnke, Clemens; Albrecht, Eva; Gieger, Christian; Khademi, Mohsen; Lima Bomfim, Izaura; Fogdell-Hahn, Anna; Link, Jenny; Alfredsson, Lars; Søndergaard, Helle Bach; Hillert, Jan; Oturai, Annette B.; Hemme, Bernhard

    2014-01-01

    JC polyomavirus (JCV) carriers with a compromised immune system, such as in HIV, or subjects on immune-modulating therapies, such as anti VLA-4 therapy may develop progressive multifocal leukoencephalopathy (PML) which is a lytic infection of oligodendrocytes in the brain. Serum antibodies to JCV mark infection occur only in 50–60% of infected individuals, and high JCV-antibody titers seem to increase the risk of developing PML. We here investigated the role of human leukocyte antigen (HLA), instrumental in immune defense in JCV antibody response. Anti-JCV antibody status, as a surrogate for JCV infection, were compared to HLA class I and II alleles in 1621 Scandinavian persons with MS and 1064 population-based Swedish controls and associations were replicated in 718 German persons with MS. HLA-alleles were determined by SNP imputation, sequence specific (SSP) kits and a reverse PCR sequence-specific oligonucleotide (PCR-SSO) method. An initial GWAS screen displayed a strong HLA class II region signal. The HLA-DRB1*15 haplotype was strongly negatively associated to JCV sero-status in Scandinavian MS cases (OR = 0.42, p = 7×10−15) and controls (OR = 0.53, p = 2×10−5). In contrast, the DQB1*06:03 haplotype was positively associated with JCV sero-status, in Scandinavian MS cases (OR = 1.63, p = 0.006), and controls (OR = 2.69, p = 1×10−5). The German dataset confirmed these findings (OR = 0.54, p = 1×10−4 and OR = 1.58, p = 0.03 respectively for these haplotypes). HLA class II restricted immune responses, and hence CD4+ T cell immunity is pivotal for JCV infection control. Alleles within the HLA-DR1*15 haplotype are associated with a protective effect on JCV infection. Alleles within the DQB1*06:03 haplotype show an opposite association. These associations between JC virus antibody response and human leucocyte antigens supports the notion that CD4+ T cells are crucial in the immune defence to JCV and lays

  5. Human Reliability Program Workshop

    SciTech Connect

    Landers, John; Rogers, Erin; Gerke, Gretchen

    2014-05-18

    A Human Reliability Program (HRP) is designed to protect national security as well as worker and public safety by continuously evaluating the reliability of those who have access to sensitive materials, facilities, and programs. Some elements of a site HRP include systematic (1) supervisory reviews, (2) medical and psychological assessments, (3) management evaluations, (4) personnel security reviews, and (4) training of HRP staff and critical positions. Over the years of implementing an HRP, the Department of Energy (DOE) has faced various challenges and overcome obstacles. During this 4-day activity, participants will examine programs that mitigate threats to nuclear security and the insider threat to include HRP, Nuclear Security Culture (NSC) Enhancement, and Employee Assistance Programs. The focus will be to develop an understanding of the need for a systematic HRP and to discuss challenges and best practices associated with mitigating the insider threat.

  6. Distribution of human polyomaviruses, adenoviruses, and hepatitis E virus in the environment and in a drinking-water treatment plant.

    PubMed

    Albinana-Gimenez, Nestor; Clemente-Casares, Pilar; Bofill-Mas, Silvia; Hundesa, Ayalkibet; Ribas, Ferran; Girones, Rosina

    2006-12-01

    Large numbers of viruses are excreted in human feces and urine, which even at low concentrations may cause illness when ingested. Some of these viruses have not been traditionally monitored in terms of waterborne diseases and are considered emergent viruses, such as hepatitis E virus (HEV) and JC and BK polyomavirus (JCPyV and BKPyV). The high prevalence of human adenoviruses (HAdV) and polyomaviruses, which both show DNA genomes, in sewage from widely divergent areas has suggested the relevance of evaluating these viruses as possible indicators of viral contamination. The concentration of these viruses was analyzed in sewage and river water and after treatment in a drinking-water treatment plant including chlorination, flocculation, ozonation, and granulate active carbon (GAC) filtration. Samples of GAC-filtered water were collected before a second chlorination treatment. The river used as a source of fresh water presented an average concentration of 2.6 x 10(1) JCPyV and 4 x 10(2) HAdV GC (genome copies)/L. A removal of 2 logarithms (99%) of HAdV and JCPyV was observed in the drinking-water treatment plant. All the GAC-filtered water samples studied contained HAdV, with a mean value of 4.3 HAdV GC/L. HEV strains belonging to genotype 3 were frequently detected in low concentrations in urban sewage and in biosolids or sewage containing swine feces but not in the river water samples studied. The detection of viruses by molecular techniques is useful for genetically describe emergent viruses in community wastewaters and water supplies. Quantification of JCPyV and HAdV using quantitative real-time PCR (QPCR) may be useful for evaluating virus removal efficiency in water treatment plants and as an index of the virological quality of water and of the potential presence of human viruses.

  7. A Retrograde Trafficking Inhibitor of Ricin and Shiga-Like Toxins Inhibits Infection of Cells by Human and Monkey Polyomaviruses

    PubMed Central

    Nelson, Christian D. S.; Carney, Dan W.; Derdowski, Aaron; Lipovsky, Alex; Gee, Gretchen V.; O’Hara, Bethany; Williard, Paul; DiMaio, Daniel; Sello, Jason K.; Atwood, Walter J.

    2013-01-01

    ABSTRACT Polyomaviruses are ubiquitous pathogens that cause severe disease in immunocompromised individuals. JC polyomavirus (JCPyV) is the causative agent of the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML), whereas BK polyomavirus (BKPyV) causes polyomavirus-induced nephropathy and hemorrhagic cystitis. Vaccines or antiviral therapies targeting these viruses do not exist, and treatments focus on reducing the underlying causes of immunosuppression. We demonstrate that retro-2cycl, an inhibitor of ricin and Shiga-like toxins (SLTs), inhibits infection by JCPyV, BKPyV, and simian virus 40. Retro-2cycl inhibits retrograde transport of polyomaviruses to the endoplasmic reticulum, a step necessary for productive infection. Retro-2cycl likely inhibits polyomaviruses in a way similar to its ricin and SLT inhibition, suggesting an overlap in the cellular host factors used by bacterial toxins and polyomaviruses. This work establishes retro-2cycl as a potential antiviral therapy that broadly inhibits polyomaviruses and possibly other pathogens that use retrograde trafficking. PMID:24222489

  8. JC Polyomavirus Infection of Primary Human Renal Epithelial Cells Is Controlled by a Type I IFN-Induced Response

    PubMed Central

    Assetta, Benedetta; De Cecco, Marco; O’Hara, Bethany

    2016-01-01

    ABSTRACT The JC and BK human polyomaviruses (JCPyV and BKPyV, respectively) establish lifelong persistent infections in the kidney. In immunosuppressed individuals, JCPyV causes progressive multifocal leukoencephalopathy (PML), a fatal neurodegenerative disease, and BKPyV causes polyomavirus-associated nephropathy (PVN). In this study, we compared JCPyV and BKPyV infections in primary human renal proximal tubule epithelial (HRPTE) cells. JCPyV established a persistent infection, but BKPyV killed the cells in 15 days. To identify the cellular factors responsible for controlling JCPyV infection and promoting viral persistence, we profiled the transcriptomes of JCPyV- and BKPyV-infected cells at several time points postinfection. We found that infection with both viruses induced interferon production but that interferon-stimulated genes (ISGs) were only activated in the JCPyV-infected cells. Phosphorylated STAT1 and IRF9, which are responsible for inducing ISGs, translocated to the nucleus of JCPyV-infected cells but did not in BKPyV-infected cells. In BKPyV-infected cells, two critical suppressors of cytokine signaling, SOCS3 and SOCS1, were induced. Infection with BKPyV but not JCPyV caused reorganization of PML bodies that are associated with inactivating antiviral responses. Blockade of the interferon receptor and neutralization of soluble interferon alpha (IFN-α) and IFN-β partially alleviated the block to JCPyV infection, leading to enhanced infectivity. Our results show that a type I IFN response contributes to the establishment of persistent infection by JCPyV in HRPTE cells. PMID:27381292

  9. Evaluating sewage-associated JCV and BKV polyomaviruses for sourcing human fecal pollution in a coastal river in Southeast Queensland, Australia.

    PubMed

    Ahmed, W; Wan, C; Goonetilleke, A; Gardner, T

    2010-01-01

    In this study, the host-sensitivity and host-specificity of JC virus (JCV) and BK virus (BKV) polyomaviruses were evaluated by testing wastewater and fecal samples from nine host groups in Southeast Queensland, Australia. The JCV and BKV polyomaviruses were detected in 63 human wastewater samples collected from primary and secondary effluent, suggesting high sensitivity of these viruses in human wastewater. In the 81 animal wastewater and fecal samples tested, 80 were polymerase chain reaction (PCR) negative for the JCV and BKV markers. Only one sample (out of 81 animal wastewater and fecal samples) from pig wastewater was positive. Nonetheless, the overall host-specificity of these viruses to differentiate between human and animal wastewater and fecal samples was 0.99. To our knowledge, this is the first study in Australia that reports on the high specificity of JCV and BKV polyomaviruses. To evaluate the field application of these viral markers for detecting human fecal pollution, 20 environmental samples were collected from a coastal river. In the 20 samples tested, 15% (3/20) and 70% (14/20) samples exceeded the regulatory guidelines for Escherichia coli and enterococci levels for marine waters. In all, five (25%) samples were PCR positive for JCV and BKV, indicating the presence of human fecal pollution in the coastal river investigated. The results suggest that JCV and BKV detection using PCR could be a useful tool for identifying human-sourced fecal pollution in coastal waters.

  10. MW polyomavirus and STL polyomavirus present in tonsillar tissues from children with chronic tonsillar disease.

    PubMed

    Peng, J; Li, K; Zhang, C; Jin, Q

    2016-01-01

    We aimed to explore the frequency of all 13 human polyomaviruses (HPyVs), especially MW polyomavirus (MWPyV) and STL polyomavirus (STLPyV), in tonsillar tissues from Chinese children with chronic tonsillar disease. We examined 99 swabs from mucosal surfaces of palatine tonsils, in which six HPyVs were detected. MWPyV and STLPyV were each detected in two samples. This provides new evidence for the hypothesis that the lymphoid system may play a role in HPyV infection and persistence. We need to define their role in tonsillar disease in the future.

  11. No association between Birt-Hogg-Dubé syndrome skin fibrofolliculomas and the first 10 described human polyomaviruses or human papillomaviruses.

    PubMed

    Bradley, Maria; Nordfors, Cecilia; Vlastos, Andrea; Ferrara, Giovanni; Ramqvist, Torbjörn; Dalianis, Tina

    2014-11-01

    The rare autosomal dominant condition Birt-Hogg-Dubé syndrome (BHD) is attributed to mutations on chromosome 17 in the folliculin (FLCN) gene, but not always diagnosed due to lack of, or a variety of symptoms such as fibrofolliculomas, lung cystic lesions, spontaneous pneumothorax and renal cancer. We hypothesized that the lack of or variability in symptoms could be due to BHD patients potentially being abnormally susceptible to infections with human papillomavirus (HPV) or human polyomavirus (HPyV), which can be associated with skin lesions or latency in the kidneys. Seven fibrofolliculoma skin lesions, one renal cancer and one lung cyst from nine patients with BHD treated at the Karolinska University Hospital were therefore analyzed for cutaneous and mucosal HPV types and 10 HPyVs by bead based multiplex assays or by PCR. All samples were negative for viral DNA. In conclusion, the data suggest that HPV and HPyVs do not contribute to BHD pathology.

  12. A Workshop in Human Sexuality for Parents

    ERIC Educational Resources Information Center

    Barnum, Barbara; Megenity, Jack

    1973-01-01

    By having workshop participants involved in verbalizing, valuing, and decision-making processes concerning human sexuality, the author feels that the parents of pre-teenagers and teenagers, are better able to understand the scope and focus of sex education in schools and churches. (GB)

  13. Archetypal and rearranged sequences of human polyomavirus JC transcription control region in peripheral blood leukocytes and in cerebrospinal fluid.

    PubMed

    Ciappi, S; Azzi, A; De Santis, R; Leoncini, F; Sterrantino, G; Mazzotta, F; Mecocci, L

    1999-04-01

    Two forms of human polyomavirus JC (JCV) genome are known based upon the structure of the transcriptional control region (TCR) of the virus: the archetypal form, which is commonly detected in urine, and the rearranged form, which was first detected in brain tissue from progressive multifocal leukoencephalopathy (PML) patients. The latter actually includes a group of TCR variants that, relative to the former, are characterized by various deletions and/or duplications. The aim of this study was to establish whether or not a correlation exists among the TCR type, the spreading of the virus within the host and its ability to cause PML. JCV TCR sequences from peripheral blood leukocytes (PBL) and cerebrospinal fluid (CSF) obtained from various groups of patients were compared. JCV with archetypal TCR was detected in CSF and PBL specimens from patients without neurological disorders or who eventually received a diagnosis of a non-PML neurological disorder. Rearranged TCR sequences were detected in all the CSF and PBL specimens from PML patients. The high similarity observed between the TCR structure detected in PBL and CSF specimens from individual patients could strengthen the hypothesis that PBL has a role in spreading JCV to the brain. Moreover, heterogeneous TCR patterns have been shown in individual PBL specimens from PML patients. This supports the hypothesis that, in PBL, JCV may replicate and undergo rearrangements of the TCR. The detection of JCV DNA by PCR in CSF independently from PML, although rare, could suggest that this assay is not sufficient for a virological diagnosis of PML. Further studies are required to assess the usefulness of quantitative assays or TCR typing in combination with PCR for diagnostic purposes.

  14. Polycyclic Aromatic Hydrocarbons—Induced ROS Accumulation Enhances Mutagenic Potential of T-Antigen From Human Polyomavirus JC

    PubMed Central

    WILK, ANNA; RSKI, PIOTR WALIGÓ; LASSAK, ADAM; VASHISTHA, HIMANSHU; LIRETTE, DAVID; TATE, DAVID; ZEA, ARNOLD H.; KOOCHEKPOUR, SHAHRIAR; RODRIGUEZ, PAULO; MEGGS, LEONARD G.; ESTRADA, JOHN J.; OCHOA, AUGUSTO; REISS, KRZYSZTOF

    2014-01-01

    Polycyclic aromatic hydrocarbons (PAHs) are the products of incomplete combustion of organic materials, which are present in cigarette smoke, deep-fried food, and in natural crude oil. Since PAH-metabolites form DNA adducts and cause oxidative DNA damage, we asked if these environmental carcinogens could affect transforming potential of the human Polyomavirus JC oncoprotein, T-antigen (JCV T-antigen). We extracted DMSO soluble PAHs from Deepwater Horizon oil spill in the Gulf of Mexico (oil-PAHs), and detected several carcinogenic PAHs. The oil-PAHs were tested in exponentially growing cultures of normal mouse fibroblasts (R508), and in R508 stably expressing JCV T-antigen (R508/T). The oil-PAHs were cytotoxic only at relatively high doses (1:50–1:100 dilution), and at 1:500 dilution the growth and cell survival rates were practically unaffected. This non-toxic dose triggered however, a significant accumulation of reactive oxygen species (ROS), caused oxidative DNA damage and the formation of DNA double strand breaks (DSBs). Although oil-PAHs induced similar levels of DNA damage in R508 and R508/T cells, only T-antigen expressing cells demonstrated inhibition of high fidelity DNA repair by homologous recombination (HRR). In contrast, low-fidelity repair by non-homologous end joining (NHEJ) was unaffected. This potential mutagenic shift between DNA repair mechanisms was accompanied by a significant increase in clonal growth of R508/T cells chronically exposed to low doses of the oil-PAHs. Our results indicate for the first time carcinogenic synergy in which oil-PAHs trigger oxidative DNA damage and JCV T-antigen compromises DNA repair fidelity. PMID:23558788

  15. Polycyclic aromatic hydrocarbons-induced ROS accumulation enhances mutagenic potential of T-antigen from human polyomavirus JC.

    PubMed

    Wilk, Anna; Waligórski, Piotr; Lassak, Adam; Vashistha, Himanshu; Lirette, David; Tate, David; Zea, Arnold H; Koochekpour, Shahriar; Rodriguez, Paulo; Meggs, Leonard G; Estrada, John J; Ochoa, Augusto; Reiss, Krzysztof

    2013-11-01

    Polycyclic aromatic hydrocarbons (PAHs) are the products of incomplete combustion of organic materials, which are present in cigarette smoke, deep-fried food, and in natural crude oil. Since PAH-metabolites form DNA adducts and cause oxidative DNA damage, we asked if these environmental carcinogens could affect transforming potential of the human Polyomavirus JC oncoprotein, T-antigen (JCV T-antigen). We extracted DMSO soluble PAHs from Deepwater Horizon oil spill in the Gulf of Mexico (oil-PAHs), and detected several carcinogenic PAHs. The oil-PAHs were tested in exponentially growing cultures of normal mouse fibroblasts (R508), and in R508 stably expressing JCV T-antigen (R508/T). The oil-PAHs were cytotoxic only at relatively high doses (1:50-1:100 dilution), and at 1:500 dilution the growth and cell survival rates were practically unaffected. This non-toxic dose triggered however, a significant accumulation of reactive oxygen species (ROS), caused oxidative DNA damage and the formation of DNA double strand breaks (DSBs). Although oil-PAHs induced similar levels of DNA damage in R508 and R508/T cells, only T-antigen expressing cells demonstrated inhibition of high fidelity DNA repair by homologous recombination (HRR). In contrast, low-fidelity repair by non-homologous end joining (NHEJ) was unaffected. This potential mutagenic shift between DNA repair mechanisms was accompanied by a significant increase in clonal growth of R508/T cells chronically exposed to low doses of the oil-PAHs. Our results indicate for the first time carcinogenic synergy in which oil-PAHs trigger oxidative DNA damage and JCV T-antigen compromises DNA repair fidelity.

  16. The cellular proteins which can associate specifically with polyomavirus middle T antigen in human 293 cells include the major human 70-kilodalton heat shock proteins.

    PubMed

    Pallas, D C; Morgan, W; Roberts, T M

    1989-11-01

    We compared the proteins which associate with middle T antigen (MT) of polyomavirus in human cells infected with Ad5(pymT), a recombinant adenovirus which directs the overexpression of MT, with the MT-associated proteins (MTAPs) previously identified in murine fibroblasts expressing MT. MTAPs of 27, 29, 36, and 63 kilodaltons (kDa) appeared to be fairly well conserved between the two species, as judged by comigration on two-dimensional gels. Several 61-kDa MTAP species detected in MT immunoprecipitates from both cell sources also comigrated on these gels. However, no protein comigrating precisely with the murine 85-kDa MTAP could be detected in the human cells. Furthermore, two proteins of 72 and 74 kDa associated with wild-type MT in the infected human cells but not in murine fibroblasts expressing MT. It had been previously reported for murine cells that the 70-kDa heat shock protein associates with a particular mutant MT but not with wild-type MT (G. Walter, A. Carbone, and W.J. Welch, J. Virol. 61:405-410, 1987). By the criteria of comigration on two-dimensional gels, tryptic peptide mapping, and immunoblotting, we showed that the 72- and 74-kDa proteins that associate with wild-type MT in human cells are the major human 70-kDa heat shock proteins.

  17. Common exposure to STL polyomavirus during childhood.

    PubMed

    Lim, Efrem S; Meinerz, Natalie M; Primi, Blake; Wang, David; Garcea, Robert L

    2014-09-01

    STL polyomavirus (STLPyV) was recently identified in human specimens. To determine seropositivity for STLPyV, we developed an ELISA and screened patient samples from 2 US cities (Denver, Colorado [500]; St. Louis, Missouri [419]). Overall seropositivity was 68%-70%. The age-stratified data suggest that STLPyV infection is widespread and commonly acquired during childhood.

  18. Detection and characterization of a novel polyomavirus in wild rodents.

    PubMed

    Orba, Yasuko; Kobayashi, Shintaro; Nakamura, Ichiro; Ishii, Akihiro; Hang'ombe, Bernard M; Mweene, Aaron S; Thomas, Yuka; Kimura, Takashi; Sawa, Hirofumi

    2011-04-01

    To investigate polyomavirus infection in wild rodents, we analysed DNA samples from the spleens of 100 wild rodents from Zambia using a broad-spectrum PCR-based assay. A previously unknown polyomavirus genome was identified in a sample from a multimammate mouse (Mastomys species) and the entire viral genome of 4899 bp was subsequently sequenced. This viral genome contained potential ORFs for the capsid proteins, VP1, VP2 and VP3, and early proteins, small t antigen and large T antigen. Phylogenetic analysis showed that it was a novel member of the family Polyomaviridae, and thus the virus was tentatively named mastomys polyomavirus. After transfection of the viral genome into several mammalian cell lines, transient expression of the VP1 and large T antigen proteins was confirmed by immunoblotting and immunocytochemical analyses. Comparison of large T antigen function in mastomys polyomavirus with that in rhesus monkey polyomavirus SV40 and human polyomavirus JC virus revealed that the large T antigen from mastomys polyomavirus interacted with the tumour suppressor protein pRb, but not with p53.

  19. Complete Sequence of the Smallest Polyomavirus Genome, Giant Guitarfish (Rhynchobatus djiddensis) Polyomavirus 1

    PubMed Central

    Dill, Jennifer A.

    2016-01-01

    Polyomaviruses are known to infect mammals and birds. Deep sequencing and metagenomic analysis identified the first polyomavirus from a cartilaginous fish, the giant guitarfish (Rhynchobatus djiddensis). Giant guitarfish polyomavirus 1 (GfPyV1) has typical polyomavirus genome organization, but is the smallest polyomavirus genome (3.96 kb) described to date. PMID:27198025

  20. 1988 workshop on human-machine symbiotic systems

    SciTech Connect

    Parker, L.E.; Weisbin, C.R.

    1988-01-01

    This report presents the proceedings of the 1988 Workshop on Human-Machine Symbiotic Systems. Held December 5-6, 1988 in Oak Ridge, Tennessee, the workshop served as a forum for the discussion of several critical issues in human-machine symbiosis: human-machine communication, autonomous task planning and execution monitoring for heterogeneous agents, dynamic task allocation, human-machine system architecture, and machine learning via experience and human observation.

  1. A Human Development Workshop on Cultural Identity for International Students.

    ERIC Educational Resources Information Center

    Castro-Abad, Cecilia

    To provide international students at New Jersey's Brookdale Community College with exercises on cultural awareness, a Human Development Workshop on Cultural Identity has been designed. The workshop includes exercises on language, cultural relationships, cultural identity, and styles of achieving. The program is designed to help students feel free…

  2. ENVIRONMENTAL-HUMAN HEALTH INTERCONNECTIONS: A WORKSHOP REPORT

    EPA Science Inventory

    A Pellston Workshop jointly sponsored by SETAC and SOT to discuss this topic of "Interconnections" was held in June, 2000 in Snowbird, Utah. This workshop was motivated by a deep concern shared by many human health, environmental, and social scientists for the interconnections, ...

  3. Merkel cell polyomavirus is present in common warts and carcinoma in situ of the skin.

    PubMed

    Mertz, Kirsten D; Pfaltz, Madeleine; Junt, Tobias; Schmid, Mirka; Fernandez Figueras, Maria Teresa; Pfaltz, Katrin; Barghorn, André; Kempf, Werner

    2010-10-01

    Polyomaviruses have been linked to diseases of immunosuppressed patients. We sought to determine the prevalence of Merkel cell polyomavirus in benign epithelial skin neoplasms and nonmelanoma skin cancer of immunosuppressed renal transplant recipients and long-term dialysis patients. Merkel cell polyomavirus DNA was detected by polymerase chain reaction (PCR) in 2 (10%) of 20 patients, in carcinomas in situ (Bowen's disease). In one of our patients with Merkel cell polyomavirus-positive carcinoma in situ, 9 (39.1%) of 23 skin lesions at various anatomical locations tested positive for Merkel cell polyomavirus sequences by PCR, including all of his common warts (4/4), half of his carcinoma in situ lesions (3/6), and 2 of his 3 seborrheic keratoses. In a second cohort of immunosuppressed renal transplant recipients, Merkel cell polyomavirus DNA was found in 1 (6.3%) of 16 common warts and in 2 (9.5%) of 21 carcinomas in situ. In immunocompetent individuals, Merkel cell polyomavirus DNA was found in 2 (6.7%) of 30 common warts and in 2 (8.3%) of 24 carcinomas in situ. DNA of other human polyomaviruses was not detected in any of the investigated skin neoplasms. We conclude that common warts and carcinomas in situ can be positive for Merkel cell polyomavirus in immunosuppressed as well as immunocompetent individuals. Remarkably, some of the Merkel cell polyomavirus-positive common warts did not contain human papillomavirus. Furthermore, Merkel cell polyomavirus can be found in various skin neoplasms of the same individual.

  4. Natural History of Polyomaviruses in Men: The HPV Infection in Men (HIM) Study

    PubMed Central

    Hampras, Shalaka S.; Giuliano, Anna R.; Lin, Hui-Yi; Fisher, Kate J.; Abrahamsen, Martha E.; McKay-Chopin, Sandrine; Gheit, Tarik; Tommasino, Massimo; Rollison, Dana E.

    2015-01-01

    Background. Several new polyomaviruses have been discovered in the last decade, including Merkel cell polyomavirus (MCPyV). Little is known about the natural history of the more recently discovered polyomaviruses. We estimated the incidence, prevalence, and persistence of 9 polyomaviruses (MCPyV, BK polyomavirus, KI polyomavirus, JC polyomavirus, WU polyomavirus, Human polyomavirus 6 [HPyV6], HPyV7, HPyV9, and Trichodysplasia spinulosa–associated polyomavirus) and examined factors associated with MCPyV infection in a prospective cohort of 209 men initially enrolled in the HPV Infection in Men (HIM) study. Methods. Participants enrolled at the US site of the HIM study were recruited into a substudy of cutaneous viral infections and followed for a median of 12.6 months. Eyebrow hair and normal skin swab specimens were obtained at each study visit, and the viral DNA load was measured using multiplex polymerase chain reaction. Results. MCPyV infection showed the highest prevalence (65.1% of normal skin swab specimens and 30.6% of eyebrow hair specimens), incidence (81.7 cases per 1000 person-months among normal skin swab specimens, and 24.1 cases per 1000 person-months among eyebrow hair specimens), and persistence (85.8% of normal skin swab specimens and 58.9% of eyebrow hair specimens) among all polyomaviruses examined. Age of >44 years (odds ratio [OR], 2.11; 95% confidence interval [CI], 1.03–4.33) and Hispanic race (OR, 2.64; 95% CI, 1.01–6.88) were associated with an increased prevalence of MCPyV infection in eyebrow hair and normal skin swab specimens, respectively. Conclusion. MCPyV infection is highly prevalent in adults, with age and race being predisposing factors. PMID:25387582

  5. Report of the second Human Genome Diversity workshop

    SciTech Connect

    1992-12-31

    The Second Human Genome Diversity Workshop was successfully held at Penn State University from October 29--31, 1992. The Workshop was essentially organized around 7 groups, each comprising approximately 10 participants, representing the sampling issues in different regions of the world. These groups worked independently, using a common format provided by the organizers; this was adjusted as needed by the individual groups. The Workshop began with a presentation of the mandate to the participants, and of the procedures to be followed during the workshop. Dr. Feldman presented a summary of the results from the First Workshop. He and the other organizers also presented brief comments giving their perspective on the objectives of the Second Workshop. Dr. Julia Bodmer discussed the study of European genetic diversity, especially in the context of the HLA experience there, and of plans to extend such studies in the coming years. She also discussed surveys of world HLA laboratories in regard to resources related to Human Genome Diversity. Dr. Mark Weiss discussed the relevance of nonhuman primate studies for understanding how demographic processes, such as mate exchange between local groups, affected the local dispersion of genetic variation. Primate population geneticists have some relevant experience in interpreting variation at this local level, in particular, with various DNA fingerprinting methods. This experience may be relevant to the Human Genome Diversity Project, in terms of practical and statistical issues.

  6. Multiorgan WU Polyomavirus Infection in Bone Marrow Transplant Recipient

    PubMed Central

    Siebrasse, Erica A.; Nguyen, Nang L.; Willby, Melisa J.; Erdman, Dean D.; Menegus, Marilyn A.

    2016-01-01

    WU polyomavirus (WUPyV) was detected in a bone marrow transplant recipient with severe acute respiratory distress syndrome who died in 2001. Crystalline lattices of polyomavirus-like particles were observed in the patient’s lung by electron microscopy. WUPyV was detected in the lung and other tissues by real-time quantitative PCR and identified in the lung and trachea by immunohistochemistry. A subset of WUPyV-positive cells in the lung had morphologic features of macrophages. Although the role of WUPyV as a human pathogen remains unclear, these results clearly demonstrate evidence for infection of respiratory tract tissues in this patient. PMID:26691850

  7. Multiorgan WU Polyomavirus Infection in Bone Marrow Transplant Recipient.

    PubMed

    Siebrasse, Erica A; Nguyen, Nang L; Willby, Melisa J; Erdman, Dean D; Menegus, Marilyn A; Wang, David

    2016-01-01

    WU polyomavirus (WUPyV) was detected in a bone marrow transplant recipient with severe acute respiratory distress syndrome who died in 2001. Crystalline lattices of polyomavirus-like particles were observed in the patient's lung by electron microscopy. WUPyV was detected in the lung and other tissues by real-time quantitative PCR and identified in the lung and trachea by immunohistochemistry. A subset of WUPyV-positive cells in the lung had morphologic features of macrophages. Although the role of WUPyV as a human pathogen remains unclear, these results clearly demonstrate evidence for infection of respiratory tract tissues in this patient.

  8. Planetary Protection Knowledge Gaps for Human Extraterrestrial Missions: Workshop Report

    NASA Technical Reports Server (NTRS)

    Race, Margaret S. (Editor); Johnson, James E. (Editor); Spry, James A. (Editor); Siegel, Bette; Conley, Catharine A.

    2015-01-01

    This report on Planetary Protection Knowledge Gaps for Human Extraterrestrial Missions summarizes the presentations, deliberations and findings of a workshop at NASA Ames Research Center, March 24-26, 2015, which was attended by more than 100 participants representing a diverse mix of science, engineering, technology, and policy areas. The main objective of the three-day workshop was to identify specific knowledge gaps that need to be addressed to make incremental progress towards the development of NASA Procedural Requirements (NPRs) for Planetary Protection during human missions to Mars.

  9. 76 FR 69743 - The Development and Evaluation of Human Cytomegalovirus Vaccines; Public Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-09

    ... Vaccines; Public Workshop AGENCY: Food and Drug Administration, HHS. ACTION: Notice of public workshop. The... Prevention, and the National Vaccine Program Office are announcing a public workshop entitled ``The Development and Evaluation of Human Cytomegalovirus Vaccines.'' The purpose of the public workshop is...

  10. 1988 workshop on human-machine symbiotic systems proceedings

    SciTech Connect

    Parker, L.E.; Weisbin, C.R.

    1989-01-01

    This report presents the proceedings of the 1988 Workshop on Human-Machine Symbiotic Systems. Held December 5--6, 1988, in Oak Ridge, Tennessee, the workshop served as a forum for the discussion of several critical issues in human-machine symbiosis: human-machine communication, autonomous task planning and execution monitoring for heterogeneous agents, dynamic task allocation, human-machine system architecture, and machine learning via experience and human observation. The presentation of overview papers by invited keynote speakers provided a background for the breakout session discussions in these five areas. The full powers furnished by the speakers are included in the proceedings, along with written summaries of the group discussions that report session conclusions and recommendations for future work.

  11. DOE Human Genome Program contractor-grantee workshop

    SciTech Connect

    1996-01-01

    This volume contains the proceedings for the DOE Human Genome Program`s Contractor-Grantee Workshop V held in Sante Fe, New Mexico January 28, February 1, 1996. Presentations were divided into sessions entitled Sequencing; Mapping; Informatics; Ethical, Legal, and Social Issues; and Infrastructure. Reports of individual projects described herein are separately indexed and abstracted for the database.

  12. Gamma interferon controls mouse polyomavirus infection in vivo.

    PubMed

    Wilson, Jarad J; Lin, Eugene; Pack, Christopher D; Frost, Elizabeth L; Hadley, Annette; Swimm, Alyson I; Wang, Jun; Dong, Ying; Breeden, Cynthia P; Kalman, Daniel; Newell, Kenneth A; Lukacher, Aron E

    2011-10-01

    Human polyomaviruses are associated with substantial morbidity in immunocompromised patients, including those with HIV/AIDS, recipients of bone marrow and kidney transplants, and individuals receiving immunomodulatory agents for autoimmune and inflammatory diseases. No effective antipolyomavirus agents are currently available, and no host determinants have been identified to predict susceptibility to polyomavirus-associated diseases. Using the mouse polyomavirus (MPyV) infection model, we recently demonstrated that perforin-granzyme exocytosis, tumor necrosis factor alpha (TNF-α), and Fas did not contribute to control of infection or virus-induced tumors. Gamma interferon (IFN-γ) was recently shown to inhibit replication by human BK polyomavirus in primary cultures of renal tubular epithelial cells. In this study, we provide evidence that IFN-γ is an important component of the host defense against MPyV infection and tumorigenesis. In immortalized and primary cells, IFN-γ reduces expression of MPyV proteins and impairs viral replication. Mice deficient for the IFN-γ receptor (IFN-γR(-/-)) maintain higher viral loads during MPyV infection and are susceptible to MPyV-induced tumors; this increased viral load is not associated with a defective MPyV-specific CD8(+) T cell response. Using an acute MPyV infection kidney transplant model, we further show that IFN-γR(-/-) donor kidneys harbor higher MPyV levels than donor kidneys from wild-type mice. Finally, administration of IFN-γ to persistently infected mice significantly reduces MPyV levels in multiple organs, including the kidney, a major reservoir for persistent mouse and human polyomavirus infections. These findings demonstrate that IFN-γ is an antiviral effector molecule for MPyV infection.

  13. Limited variation during circulation of a polyomavirus in the human population involves the COCO-VA toggling site of Middle and Alternative T-antigen(s).

    PubMed

    Kazem, Siamaque; Lauber, Chris; van der Meijden, Els; Kooijman, Sander; Kravchenko, Alexander A; Feltkamp, Mariet C W; Gorbalenya, Alexander E

    2016-01-01

    We have recently shown that the trichodysplasia spinulosa-associated polyomavirus (TSPyV) belongs to a large monophyletic group of mammalian polyomaviruses that experienced accelerated codon-constrained Val-Ala (COCO-VA) toggling at a protein site common to both Middle and Alternative T-antigens (MT/ALTO). Here we analyzed thirteen, mostly newly sequenced TSPyV genomes, representing ~40% of reported TS disease cases world-wide. We found two deletions and 30 variable sites (≤0.6%) that included only four sites with non-synonymous substitutions (NSS). One NSS site was under positive selection in the exon shared by Small and Middle T antigens, while three others were segregated in MT/ALTO. Two MT/ALTO sites covaried with five sites elsewhere in the genome and determined separation of twelve TSPyVs into two most populous phylogenetic lineages. The other, most distant TSPyV was distinguished by NSS at the COCO-VA site, observed for the first time during intra-species evolution. Our findings reveal a connection between micro- and macro-evolution of polyomaviruses.

  14. Mouse Polyomavirus: Propagation, Purification, Quantification, and Storage.

    PubMed

    Horníková, Lenka; Žíla, Vojtěch; Španielová, Hana; Forstová, Jitka

    2015-08-03

    Mouse polyomavirus (MPyV) is a member of the Polyomaviridae family, which comprises non-enveloped tumorigenic viruses infecting various vertebrates including humans and causing different pathogenic responses in the infected organisms. Despite the variations in host tropism and pathogenicity, the structure of the virions of these viruses is similar. The capsid, with icosahedral symmetry (ø, 45 nm, T = 7d), is composed of a shell of 72 capsomeres of structural proteins, arranged around the nucleocore containing approximately 5-kbp-long circular dsDNA in complex with cellular histones. MPyV has been one of the most studied polyomaviruses and serves as a model virus for studies of the mechanisms of cell transformation and virus trafficking, and for use in nanotechnology. It can be propagated in primary mouse cells (e.g., in whole mouse embryo cells) or in mouse epithelial or fibroblast cell lines. In this unit, propagation, purification, quantification, and storage of MPyV virions are presented.

  15. The Ancient Evolutionary History of Polyomaviruses.

    PubMed

    Buck, Christopher B; Van Doorslaer, Koenraad; Peretti, Alberto; Geoghegan, Eileen M; Tisza, Michael J; An, Ping; Katz, Joshua P; Pipas, James M; McBride, Alison A; Camus, Alvin C; McDermott, Alexa J; Dill, Jennifer A; Delwart, Eric; Ng, Terry F F; Farkas, Kata; Austin, Charlotte; Kraberger, Simona; Davison, William; Pastrana, Diana V; Varsani, Arvind

    2016-04-01

    Polyomaviruses are a family of DNA tumor viruses that are known to infect mammals and birds. To investigate the deeper evolutionary history of the family, we used a combination of viral metagenomics, bioinformatics, and structural modeling approaches to identify and characterize polyomavirus sequences associated with fish and arthropods. Analyses drawing upon the divergent new sequences indicate that polyomaviruses have been gradually co-evolving with their animal hosts for at least half a billion years. Phylogenetic analyses of individual polyomavirus genes suggest that some modern polyomavirus species arose after ancient recombination events involving distantly related polyomavirus lineages. The improved evolutionary model provides a useful platform for developing a more accurate taxonomic classification system for the viral family Polyomaviridae.

  16. The Ancient Evolutionary History of Polyomaviruses

    PubMed Central

    Buck, Christopher B.; Van Doorslaer, Koenraad; Peretti, Alberto; Geoghegan, Eileen M.; Tisza, Michael J.; An, Ping; Katz, Joshua P.; Pipas, James M.; McBride, Alison A.; Camus, Alvin C.; McDermott, Alexa J.; Dill, Jennifer A.; Delwart, Eric; Ng, Terry F. F.; Farkas, Kata; Austin, Charlotte; Kraberger, Simona; Davison, William; Pastrana, Diana V.; Varsani, Arvind

    2016-01-01

    Polyomaviruses are a family of DNA tumor viruses that are known to infect mammals and birds. To investigate the deeper evolutionary history of the family, we used a combination of viral metagenomics, bioinformatics, and structural modeling approaches to identify and characterize polyomavirus sequences associated with fish and arthropods. Analyses drawing upon the divergent new sequences indicate that polyomaviruses have been gradually co-evolving with their animal hosts for at least half a billion years. Phylogenetic analyses of individual polyomavirus genes suggest that some modern polyomavirus species arose after ancient recombination events involving distantly related polyomavirus lineages. The improved evolutionary model provides a useful platform for developing a more accurate taxonomic classification system for the viral family Polyomaviridae. PMID:27093155

  17. Fourth international workshop on human chromosome 5. Final progress report

    SciTech Connect

    McPherson, J.D.

    1996-12-31

    The Fourth International Workshop on Human Chromosome 5 was held in Manchester, UK on November 9--10, 1996 and was hosted by the University of Manchester. The major goals of the workshop were: (1) to collate the various genetic, cytogenetic and physical maps of human chromosome 5; (2) to integrate these maps and identify/correct discrepancies between them wherever possible; (3) to catalogue the sequence-ready contigs of the chromosome; (4) to co-ordinate the various sequencing efforts to avoid future duplication; (5) to establish the first (to the author`s knowledge) web site for the human chromosome 5 community which contains the above information in a readily accessible form.

  18. DOE/FDA/EPA: Workshop on methylmercury and human health

    SciTech Connect

    Moskowitz, P.D.; Saroff, L.; Bolger, M.; Cicmanec, J.; Durkee, S.

    1994-12-31

    In the US the general population is exposed to methylmercury (MeHg) principally through the consumption of fish. There is continuing discussion about the sources of this form of mercury (Hg), the magnitudes and trends in exposures to consumers, and the significance of the sources and their contributions to human health. In response to these discussions, the US Department of Energy, the US Food and Drug Administration, and the US Environmental Protection Agency cosponsored a two-day workshop to discuss data and methods available for characterizing the risk to human health presented by MeHg. This workshop was attended by 45 individuals representing various Federal and state organizations and interested stakeholders. The agenda covered: Agency interests; probabilistic approach to risk assessment; emission sources; atmospheric transport; biogeochemical cycling; exposure assessment; health effects of MeHg; and research needs.

  19. Polyomavirus T Antigens Activate an Antiviral State

    PubMed Central

    Giacobbi, Nicholas S.; Gupta, Tushar; Coxon, Andrew; Pipas, James M.

    2014-01-01

    Ectopic expression of Simian Virus 40 (SV40) large T antigen (LT) in mouse embryonic fibroblasts (MEFs) increased levels of mRNAs encoding interferon stimulated genes (ISGs). The mechanism by which T antigen increases levels of ISGs in MEFs remains unclear. We present evidence that expression of T antigen from SV40, Human Polyomaviruses BK (BKV) or JC (JCV) upregulate production of ISGs in MEFs, and subsequently result in an antiviral state, as determined by inhibition of VSV or EMCV growth. The first 136 amino acids of LT are sufficient for these activities. Furthermore, increased ISG expression and induction of the antiviral state requires STAT1. Finally, the RB binding motif of LT is necessary for activation of STAT1. We conclude that the induction of the STAT1 mediated innate immune response in MEFs is a common feature shared by SV40, BKV and JCV. PMID:25589241

  20. Seroprevalence of trichodysplasia spinulosa-associated polyomavirus.

    PubMed

    van der Meijden, Els; Kazem, Siamaque; Burgers, Manda M; Janssens, Rene; Bouwes Bavinck, Jan Nico; de Melker, Hester; Feltkamp, Mariet C W

    2011-08-01

    We identified a new polyomavirus in skin lesions from a patient with trichodysplasia spinulosa (TS). Apart from TS being an extremely rare disease, little is known of its epidemiology. On the basis of knowledge regarding other polyomaviruses, we anticipated that infections with trichodysplasia spinulosa-associated polyomavirus (TSV) occur frequently and become symptomatic only in immunocompromised patients. To investigate this hypothesis, we developed and used a Luminex-based TSV viral protein 1 immunoassay, excluded cross-reactivity with phylogenetically related Merkel cell polyomavirus, and measured TSV seroreactivity. Highest reactivity was found in a TS patient. In 528 healthy persons in the Netherlands, a wide range of seroreactivities was measured and resulted in an overall TSV seroprevalence of 70% (range 10% in small children to 80% in adults). In 80 renal transplant patients, seroprevalence was 89%. Infection with the new TSV polyomavirus is common and occurs primarily at a young age.

  1. Polyomaviruses KI and WU in immunocompromised patients with respiratory disease.

    PubMed

    Mourez, Thomas; Bergeron, Anne; Ribaud, Patricia; Scieux, Catherine; de Latour, Régis Peffault; Tazi, Abdellatif; Socié, Gérard; Simon, François; LeGoff, Jérôme

    2009-01-01

    Polyomaviruses KI (KIPyV) and WU (WUPyV) were recently identified, mainly in respiratory specimens from children. Among 200 patients with respiratory disorders admitted to Saint Louis Hospital, Paris, France, KIPyV was detected in 8% and WUPyV in 1%. KIPyV was significantly more frequent among human stem cell transplant patients (17.8% vs. 5.1%; p = 0.01).

  2. Polyomaviruses KI and WU in Immunocompromised Patients with Respiratory Disease

    PubMed Central

    Mourez, Thomas; Bergeron, Anne; Ribaud, Patricia; Scieux, Catherine; Peffault de Latour, Régis; Tazi, Abdellatif; Socié, Gérard; Simon, François

    2009-01-01

    Polyomaviruses KI (KIPyV) and WU (WUPyV) were recently identified, mainly in respiratory specimens from children. Among 200 patients with respiratory disorders admitted to Saint Louis Hospital, Paris, France, KIPyV was detected in 8% and WUPyV in 1%. KIPyV was significantly more frequent among human stem cell transplant patients (17.8% vs. 5.1%; p = 0.01). PMID:19116066

  3. The third international workshop of human chromosome 5. Final report

    SciTech Connect

    1994-12-31

    The Third International Workshop on Human Chromosome 5 was held in Laguna Beach, California, March 5-8, 1994. The pace at which new mapping information has been published in the last year make almost any report outdated before publication. Much of the information in this report and the most recent data from the Human chromosome 5 Genome Center at U.C. Irvine on the physical map of chromosome 5 are accessible via a WWW server. For most loci referred to in this report that can be detected by Polymerase Chain Reaction, the sequences of the oligonucleotide primers are available and some primer sequences are provided in this report.

  4. Occurrence, genotypic characterization, and patterns of shedding of human polyomavirus JCPyV and BKPyV in urine samples of healthy individuals in São Paulo, Brazil.

    PubMed

    Urbano, Paulo Roberto Palma; Oliveira, Renato Reis; Romano, Camila Malta; Pannuti, Claudio Sergio; Fink, Maria Cristina Domingues da Silva

    2016-01-01

    The objective of this study was to evaluate the prevalence, genotypic characterization, and determination of the patterns of shedding of human polyomavirus JC (JCPyV) and BK (BKPyV) in consecutive urine samples collected from healthy adults. Urine samples collected monthly over a 6 month period were screened by polymerase chain reaction (PCR) with two sets of primers complementary to the VP1 protein region specific for the JCPyV or BKPyV genome. The viral load of JCPyV and BKPyV in positive samples was determined by quantitative real time PCR. Seventy-one healthy individuals (ages between 18 and 65) were included in the study. Polyomavirus DNA urinary shedding was identified in 44 (62%) of the 71 individuals evaluated: BKPyV only in 16 (22.5%); JCPyV only in 19 (26.7%); and both in 9 (12.7%). Among the 28 individuals shedding JCPyV, the shedding was nearly continuous in 13 (46.4%) and sporadic in 15 (53.6%), whereas all BKPyV shedding was sporadic. A total of 45 (19 BKPyV and 26 JCPyV) strains were identified. Of the BKPyV strains, individuals were observed that excreted all genotypes except genotype 3 and the JCPyV strains, excretion of 5 different genotypes. Evaluating the age of individuals who excrete JCPyV and BKPyV, mostly are young adults, with a slight increase with increasing age and observing the viral load can not draw any parallel between the increase or decrease of age or excreted genotype as there was a wide variation both in the excretion of BKPyV and JCPyV. The high occurrence of isolated or simultaneous urinary shedding of JCPyV and BKPyV in healthy individuals merits further study.

  5. Workshop on Science and the Human Exploration of Mars

    NASA Technical Reports Server (NTRS)

    Duke, M. B. (Editor)

    2001-01-01

    The exploration of Mars will be a multi-decadal activity. Currently, a scientific program is underway, sponsored by NASA's Office of Space Science in the United States, in collaboration with international partners France, Italy, and the European Space Agency. Plans exist for the continuation of this robotic program through the first automated return of Martian samples in 2014. Mars is also a prime long-term objective for human exploration, and within NASA, efforts are being made to provide the best integration of the robotic program and future human exploration missions. From the perspective of human exploration missions, it is important to understand the scientific objectives of human missions, in order to design the appropriate systems, tools, and operational capabilities to maximize science on those missions. In addition, data from the robotic missions can provide critical environmental data - surface morphology, materials composition, evaluations of potential toxicity of surface materials, radiation, electrical and other physical properties of the Martian environment, and assessments of the probability that humans would encounter Martian life forms. Understanding of the data needs can lead to the definition of experiments that can be done in the near-term that will make the design of human missions more effective. This workshop was convened to begin a dialog between the scientific community that is central to the robotic exploration mission program and a set of experts in systems and technologies that are critical to human exploration missions. The charge to the workshop was to develop an understanding of the types of scientific exploration that would be best suited to the human exploration missions and the capabilities and limitations of human explorers in undertaking science on those missions.

  6. Exposure to raccoon polyomavirus (RacPyV) in free-ranging North American raccoons (Procyon lotor)

    PubMed Central

    Cruz Dela, FN; Estrada, M; Leutenegger, CM; Pesavento, PA; Woolard, KD

    2016-01-01

    There is evidence that raccoon polyomavirus is causative for neuroglial brain tumors in the western United States. It is unknown if infection is limited to geographic locales where tumors have been reported or is widespread, like human polyomaviruses. We demonstrate raccoons in western, eastern and midwestern states have been exposed to RacPyV by detection of antibodies to capsid protein, VP1. While raccoons in eastern and midwestern states are seropositive, exposure is lower than in the western states. Additionally, across geographic areas seropositivity is higher in older compared to younger raccoons, similar to polyomavirus exposure in humans. Serum titers are significantly higher in raccoons with tumors compared to raccoons without. Unlike polyomavirus-associated diseases in humans, we did not detect significant sequence variation between tumor and non-tumor tissue in raccoons with tumors compared to those without tumors. This warrants further investigation into co-morbid diseases or genetic susceptibility studies of the host. PMID:26802526

  7. Exposure to raccoon polyomavirus (RacPyV) in free-ranging North American raccoons (Procyon lotor).

    PubMed

    Church, M E; Dela Cruz, F N; Estrada, M; Leutenegger, C M; Pesavento, P A; Woolard, K D

    2016-02-01

    There is evidence that raccoon polyomavirus is causative for neuroglial brain tumors in the western United States. It is unknown if infection is limited to geographic locales where tumors have been reported or is widespread, like human polyomaviruses. We demonstrate raccoons in western, eastern and midwestern states have been exposed to RacPyV by detection of antibodies to capsid protein, VP1. While raccoons in eastern and midwestern states are seropositive, exposure is lower than in the western states. Additionally, across geographic areas seropositivity is higher in older as compared to younger raccoons, similar to polyomavirus exposure in humans. Serum titers are significantly higher in raccoons with tumors compared to raccoons without. Unlike polyomavirus-associated diseases in humans, we did not detect significant sequence variation between tumor and non-tumor tissue in raccoons with tumors compared to those without tumors. This warrants further investigation into co-morbid diseases or genetic susceptibility studies of the host.

  8. A molecular case-control study of the Merkel cell polyomavirus in colon cancer.

    PubMed

    Campello, Cesare; Comar, Manola; D'Agaro, Pierlanfranco; Minicozzi, Anna; Rodella, Luca; Poli, Albino

    2011-04-01

    To explore the putative role of the Merkel cell polyomavirus in human colon cancer, a prospective molecular case-control study was undertaken in patients and their relatives enrolled during a screening program. Fresh tissue samples from 64 cases of colon cancer (mean age 69.9 ± 11.0 years; 40 males) and fresh biopsies from 80 relatives (mean age 53.7 ± 8.6 years; 43 male; 55 son/daughter, 23 brother/sister, 2 parents) were analyzed by PCR and sequencing. Pre-cancerous lesions, namely adenomas and polyps, were detected in 15 (18.8%) and 9 (11.2%) of the controls, respectively. In addition, 144 blood samples were examined. Merkel cell polyomavirus DNA was detected in 6.3% of cases and 8.8% of controls. This difference was not statistically significant in the logistic regression analysis, after adjustment for age. Whereas blood samples from both cases and controls tested negative, the DNA Merkel cell polyomavirus was identified in 12.5% of adenoma/polyp tissues. No statistically significant difference was found when prevalence rates of Merkel cell polyomavirus in normal, pre-cancerous and cancer tissues were compared. Sequence analysis of the viral LT3 and VP1 regions showed high homology (>99%) with those of strains circulating worldwide, especially with genotypes detected in France. The findings of this survey are consistent with the hypothesis that the Merkel cell polyomavirus, in addition to other human polyomaviruses, can be recovered frequently from the gastrointestinal tract, because it is transmitted throughout the fecal-oral route. Moreover, the study does not indicate a role for Merkel cell polyomavirus in the genesis of colon cancer.

  9. Report on the Second International Workshop on Human Chromosome 9

    SciTech Connect

    Kwiatkowski, D.J.; Armour, J.; Bale, A.E.

    1993-12-31

    The Second International Workshop on Human Chromosome 9 was held in Chatham, Massachusetts on April 18--20, 1993. Fifty-three abstracts were received and the data presented on posters. The purpose of the meeting was to bring together all interested investigators working on the map of chromosome 9, many of whom had disease-specific interests. After a brief presentation of interests and highlighted results, the meeting broke up into the following subgroups for production of consensus maps: 9p; 9cen-q32; 9q32 ter. A global mapping group also met. Reports of each of these working groups is presented in the summary.

  10. The Delivery of Human Services. Report of the Workshop (New York, New York, November 23-25, 1986). Youth Corps Exchange Project 1986-1987 Workshops.

    ERIC Educational Resources Information Center

    Jucovy, Linda

    This workshop report deals with developing a youth service corps. Twenty-nine representatives from 15 youth corps attended the workshop, the second in a series of six sponsored by the Youth Corps Exchange Project. The report begins with an overview of the highlights of the workshop. The second section, which discusses the need for human services,…

  11. Genotypic diversity of polyomaviruses circulating among kidney transplant recipients in Kuwait.

    PubMed

    Chehadeh, Wassim; Nampoory, Mangalathillam Raman

    2013-09-01

    BK virus (BKV) and JC virus (JCV) are human polyomaviruses that cause asymptomatic latent infections. Under immunosuppression, BKV-associated nephropathy has been documented in Kuwait and elsewhere. Even though different BKV and JCV genotypes with distinct geographical distribution have been described, the genotype of polyomavirus detected in Kuwait is still unknown. The aim of this study was to determine the genotypes of BKV and JCV detected in renal transplant recipients. The detection of polyomavirus DNA was carried out in serum and urine samples of 200 post-transplant recipients during a 1-year follow-up period. Fifty-one (25.5%) post-transplant recipients were tested positive for polyomavirus DNA by semi-nested PCR. JCV DNA could be detected in 29 (57%) patients, and BKV DNA in 22 (43%) patients. In two renal transplant recipients, both BKV and JCV were detected. According to the Bayesian phylogenetic analysis of polyomavirus VP1 sequences, the majority of detected BKV sequences were most closely related to genotypes I and IV, whereas the majority of JCV sequences were most closely related to genotype 3. Polyomavirus VP1 sequences showed strong stability for up to 12 months in most patients; however, in one patient, an amino acid substitution in the BKV VP1 protein was identified over time. The results suggest a close relationship of BKV sequences with the Asian and European strains, and of JCV sequences with the African strains. Long follow-up studies are needed to investigate the association of polyomavirus polymorphism or genotypic shift with the development of nephropathy.

  12. Complete Genome Sequence of a Novel WU Polyomavirus Isolate from Arkansas, USA, Associated with Acute Respiratory Infection

    PubMed Central

    Denson, J. L.; Schwalm, K. S.; Stoner, A. N.; Kincaid, J. C.; Abramo, T. J.; Thompson, T. M.; Ulloa, E. M.; Burchiel, S. W.; Dinwiddie, D. L.

    2017-01-01

    ABSTRACT We report here the complete genome sequence of a WU polyomavirus (WUPyV) isolate, also known as human polyomavirus 4, collected in 2016 from a patient in Arkansas with an acute respiratory infection. Isolate hPyV4/USA/AR001/2016 has a double-stranded DNA genome of 5,229 bp in length. PMID:28082496

  13. Review of the 1988 workshop on human-machine symbiotic systems

    SciTech Connect

    Parker, L.E.; Weisbin, C.R.

    1989-01-01

    This report presents a review of the 1988 Workshop on Human-Machine Symbiotic Systems. Held December 5--6, 1988 in Oak Ridge, Tennessee, the workshop served as a forum for the discussion of several critical issues in human-machine symbiosis: human-machine communication, autonomous task planning and execution monitoring for heterogeneous agents, dynamic task allocation, human-machine system architecture, and machine learning via experience and human observation. The presentation of overview papers by invited keynote speakers provided a background for the breakout session discussions in these five areas. A summary of the conclusions and recommendations for future work resulting from the workshop is reported. 6 refs.

  14. BK polyomavirus association with colorectal cancer development.

    PubMed

    Khabaz, M N; Nedjadi, T; Gari, M A; Al-Maghrabi, J A; Atta, H M; Basuni, A A; Elderwi, D A

    2016-05-06

    The development of human neoplasms can be provoked by exposure to one of several viruses. Burkitt lymphoma, cervical carcinoma, and hepatocellular carcinoma are associated with Epstein-Barr, human papilloma, and hepatitis B virus infections, respectively. Over the past three decades, many studies have attempted to establish an association between colorectal cancer and viruses, with debatable results. The aim of the present research was to assess the presence of BK polyomavirus (BKV) DNA and protein in colorectal cancer samples from patients in the Western Province of Saudi Arabia. DNA extracted from archival samples of colorectal cancer tissues was analyzed for BKV sequences using polymerase chain reaction (PCR)-based techniques. In addition, expression of a BKV protein was assessed using immunohistochemical staining. None of the tumor and control samples examined tested positive for BKV DNA in PCR assays. Furthermore, immunohistochemical staining failed to detect viral proteins in both cancer and control specimens. These results may indicate that BKV is not associated with the development of colorectal adenocarcinoma in patients in the Western Province of Saudi Arabia.

  15. Quantitative Proteomic Analysis of Enriched Nuclear Fractions from BK Polyomavirus-infected Primary Renal Proximal Tubule Epithelial Cells

    PubMed Central

    Justice, Joshua L.; Verhalen, Brandy; Kumar, Ranjit; Lefkowitz, Elliot J.; Imperiale, Michael J.; Jiang, Mengxi

    2016-01-01

    Polyomaviruses are a family of small DNA viruses that are associated with a number of severe human diseases, particularly in immunocompromised individuals. The detailed virus-host interactions during lytic polyomavirus infection are not fully understood. Here we report the first nuclear proteomic study with BK polyomavirus (BKPyV) in a primary renal proximal tubule epithelial cell culture system using stable isotope labeling by amino acids in cell culture (SILAC) proteomic profiling coupled with LC-MS/MS. We demonstrated the feasibility of SILAC labeling in these primary cells and subsequently performed reciprocal labeling-infection experiments to identify proteins that are altered by BKPyV infection. Our analyses revealed specific proteins that are significantly up- or down-regulated in the infected nuclear proteome. The genes encoding many of these proteins were not identified in a previous microarray study, suggesting that differential regulation of these proteins may be independent of transcriptional control. Western blotting experiments verified the SILAC proteomic findings. Finally, pathway and network analyses indicated that the host cell DNA damage response signaling and DNA repair pathways are among the cellular processes most affected at the protein level during polyomavirus infection. Our study provides a comprehensive view of the host nuclear proteomic changes during polyomavirus lytic infection and suggests potential novel host factors required for a productive polyomavirus infection. PMID:26354146

  16. A Human Sexuality Workshop for Community Professionals: Its Development and Evaluation.

    ERIC Educational Resources Information Center

    Braverman, Barbara B.; Levine, Steven G.

    A 2-day workshop in human sexuality was planned, implemented, and evaluated by the Genesee Region Family Planning Program, Inc., Rochester, New York. The goal of the workshop was to increase participant tolerance to a variety of patterns of sexual behavior. To measure attitudinal change, the Minnesota Sex Attitude Survey was administered one month…

  17. Increased Prevalence of Human Polyomavirus JC Viruria in Chronic Inflammatory Rheumatic Diseases Patients in Treatment with Anti-TNF α: A 18 Month Follow-Up Study

    PubMed Central

    Rodio, Donatella Maria; Anzivino, Elena; Mischitelli, Monica; Bellizzi, Anna; Scrivo, Rossana; Scribano, Daniela; Conte, Gianlorenzo; Prezioso, Carla; Trancassini, Maria; Valesini, Guido; Palamara, Anna Teresa; Pietropaolo, Valeria

    2016-01-01

    Chronic inflammatory rheumatic diseases (CIRDs) are immune-mediated pathologies involving joints. To date, TNFα-blocking agents administration is the most promising therapy, although these treatments are associated with an increased Polyomavirus JC (JCPyV) reactivation, the etiological agent of the Progressive Multifocal Leukoencephalopathy (PML). The aim of this study was the recruitment and the analysis of a CIRDs cohort in order to investigate a possible correlation between JCPyV presence and the influence of anti-TNF-α agents on viral loads. Blood and urine samples were collected from 34 CIRDs subjects prior the first anti-TNF-α infusion (T0) and after 3 (T3), 6 (T6), 12 (T12), and 18 (T18) months. Results showed persistent JC viruria significantly higher than JC viremia throughout the 18 month follow-up study (p = 0.002). In JCPyV positive samples, the non-coding control region (NCCR) was analyzed. Results evidenced archetypal structures (type II-S) in all isolates with the exception of a sequence isolated from a plasma sample, that corresponds to the type II-R found in PML subjects. Finally, the viral protein 1 (VP1) genotyping was performed and results showed the prevalence of the European genotypes 1A, 1B, and 4. Since only few studies have been carried out to understand whether there is a PML risk in CIRDs population infected by JCPyV, this study contributes to enrich literature insight on JCPyV biology in this cluster. Further investigations are necessary in order to recognize the real impact of biologics on JCPyV life cycle and to identify possible and specific viral variants related to increased virulence in CIRDs patients. PMID:27242700

  18. The Raccoon Polyomavirus Genome and Tumor Antigen Transcription Are Stable and Abundant in Neuroglial Tumors

    PubMed Central

    Brostoff, Terza; Dela Cruz, Florante N.; Church, Molly E.; Woolard, Kevin D.

    2014-01-01

    ABSTRACT Raccoon polyomavirus (RacPyV) is associated with 100% of neuroglial tumors in free-ranging raccoons. Other tumor-associated polyomaviruses (PyVs), including simian virus 40 (SV40), murine PyV, and Merkel cell PyV, are found integrated in the host genome in neoplastic cells, where they constitutively express splice variants of the tumor antigen (TAg) gene. We have previously reported that RacPyV exists only as an episome (nonintegrated) in neuroglial tumors. Here, we have investigated TAg transcription in primary tumor tissue by transcriptome analysis, and we identified the alternatively spliced TAg transcripts for RacPyV. We also determined that TAg was highly transcribed relative to host cellular genes. We further colocalized TAg DNA and mRNA by in situ hybridization and found that the majority of tumor cells showed positive staining. Lastly, we examined the stability of the viral genome and TAg transcription by quantitative reverse transcriptase PCR in cultured tumor cells in vitro and in a mouse xenograft model. When tumor cells were cultured in vitro, TAg transcription increased nearly 2 log-fold over that of parental tumor tissue by passage 17. Both episomal viral genome and TAg transcription were faithfully maintained in culture and in tumors arising from xenotransplantation of cultured cells in mice. This study represents a minimal criterion for RacPyV's association with neuroglial tumors and a novel mechanism of stability for a polyomavirus in cancer. IMPORTANCE The natural cycle of polyomaviruses in mammals is to persist in the host without causing disease, but they can cause cancer in humans or in other animals. Because this is an unpredictable and rare event, the oncogenic potential of polyomavirus is primarily evaluated in laboratory animal models. Recently, raccoon polyomavirus (RacPyV) was identified in neuroglial tumors of free-ranging raccoons. Viral copy number was consistently high in these tumors but was low or undetectable in nontumor

  19. Workshop meeting report Organs-on-Chips: human disease models.

    PubMed

    van de Stolpe, Anja; den Toonder, Jaap

    2013-09-21

    The concept of "Organs-on-Chips" has recently evolved and has been described as 3D (mini-) organs or tissues consisting of multiple and different cell types interacting with each other under closely controlled conditions, grown in a microfluidic chip, and mimicking the complex structures and cellular interactions in and between different cell types and organs in vivo, enabling the real time monitoring of cellular processes. In combination with the emerging iPSC (induced pluripotent stem cell) field this development offers unprecedented opportunities to develop human in vitro models for healthy and diseased organ tissues, enabling the investigation of fundamental mechanisms in disease development, drug toxicity screening, drug target discovery and drug development, and the replacement of animal testing. Capturing the genetic background of the iPSC donor in the organ or disease model carries the promise to move towards "in vitro clinical trials", reducing costs for drug development and furthering the concept of personalized medicine and companion diagnostics. During the Lorentz workshop (Leiden, September 2012) an international multidisciplinary group of experts discussed the current state of the art, available and emerging technologies, applications and how to proceed in the field. Organ-on-a-chip platform technologies are expected to revolutionize cell biology in general and drug development in particular.

  20. Polyomavirus and Naturally Occuring Neuroglial Tumors in Raccoons (Procyon Lotor).

    PubMed

    Pesavento, Patricia A; Brostoff, Terza; Church, Molly E; Dela Cruz, Florante N; Woolard, Kevin D

    2016-01-01

    Polyomavirus (PyV) infections are widespread in human populations and, although generally associated with silent persistence, rarely cause severe disease. Among diseases convincingly associated with natural PyV infections of humans, there are remarkably different tissue tropisms and outcomes, including progressive multifocal leukoencephalopathy, transient or progressive nephropathy, and cancer. The variable character and unpredictable outcomes of infection attest to large gaps in our basic understanding of PyV biology. In particular, the rich history of research demonstrating the oncogenic potential of PyVs in laboratory animals begs the question of why cancer is not more often associated with infection. Raccoon polyomavirus (RacPyV), discovered in 2010, is consistently identified in neuroglial tumors in free-ranging raccoons in the western United States. Exposure to RacPyV is widespread, and RacPyV is detected in tissues of raccoons without tumors. Studying the relationship of RacPyV with its natural host is a unique opportunity to uncover cogent cellular targets and protein interactions between the virus and its host. Our hypothesis is that RacPyV, as an intact episome, alters cellular pathways within neural progenitor cells and drives oncogenesis.

  1. Rad51 activates polyomavirus JC early transcription.

    PubMed

    White, Martyn K; Kaminski, Rafal; Khalili, Kamel; Wollebo, Hassen S

    2014-01-01

    The human neurotropic polyomavirus JC (JCV) causes the fatal CNS demyelinating disease progressive multifocal leukoencephalopathy (PML). JCV infection is very common and after primary infection, the virus is able to persist in an asymptomatic state. Rarely, and usually only under conditions of immune impairment, JCV re-emerges to actively replicate in the astrocytes and oligodendrocytes of the brain causing PML. The regulatory events involved in the reactivation of active viral replication in PML are not well understood but previous studies have implicated the transcription factor NF-κB acting at a well-characterized site in the JCV noncoding control region (NCCR). NF-κB in turn is regulated in a number of ways including activation by cytokines such as TNF-α, interactions with other transcription factors and epigenetic events involving protein acetylation--all of which can regulate the transcriptional activity of JCV. Active JCV infection is marked by the occurrence of rapid and extensive DNA damage in the host cell and the induction of the expression of cellular proteins involved in DNA repair including Rad51, a major component of the homologous recombination-directed double-strand break DNA repair machinery. Here we show that increased Rad51 expression activates the JCV early promoter. This activation is co-operative with the stimulation caused by NF-κB p65, abrogated by mutation of the NF-κB binding site or siRNA to NFκB p65 and enhanced by the histone deacetylase inhibitor sodium butyrate. These data indicate that the induction of Rad51 resulting from infection with JCV acts through NF-κB via its binding site to stimulate JCV early transcription. We suggest that this provides a novel positive feedback mechanism to enhance viral gene expression during the early stage of JCV infection.

  2. Assessing Host-Virus Codivergence for Close Relatives of Merkel Cell Polyomavirus Infecting African Great Apes

    PubMed Central

    Madinda, Nadège F.; Ehlers, Bernhard; Wertheim, Joel O.; Akoua-Koffi, Chantal; Bergl, Richard A.; Boesch, Christophe; Akonkwa, Dieudonné Boji Mungu; Eckardt, Winnie; Fruth, Barbara; Gillespie, Thomas R.; Gray, Maryke; Hohmann, Gottfried; Karhemere, Stomy; Kujirakwinja, Deo; Langergraber, Kevin; Muyembe, Jean-Jacques; Nishuli, Radar; Pauly, Maude; Petrzelkova, Klara J.; Robbins, Martha M.; Todd, Angelique; Schubert, Grit; Stoinski, Tara S.; Wittig, Roman M.; Zuberbühler, Klaus; Peeters, Martine; Leendertz, Fabian H.

    2016-01-01

    ABSTRACT It has long been hypothesized that polyomaviruses (PyV; family Polyomaviridae) codiverged with their animal hosts. In contrast, recent analyses suggested that codivergence may only marginally influence the evolution of PyV. We reassess this question by focusing on a single lineage of PyV infecting hominine hosts, the Merkel cell polyomavirus (MCPyV) lineage. By characterizing the genetic diversity of these viruses in seven African great ape taxa, we show that they exhibit very strong host specificity. Reconciliation analyses identify more codivergence than noncodivergence events. In addition, we find that a number of host and PyV divergence events are synchronous. Collectively, our results support codivergence as the dominant process at play during the evolution of the MCPyV lineage. More generally, our results add to the growing body of evidence suggesting an ancient and stable association of PyV and their animal hosts. IMPORTANCE The processes involved in viral evolution and the interaction of viruses with their hosts are of great scientific interest and public health relevance. It has long been thought that the genetic diversity of double-stranded DNA viruses was generated over long periods of time, similar to typical host evolutionary timescales. This was also hypothesized for polyomaviruses (family Polyomaviridae), a group comprising several human pathogens, but this remains a point of controversy. Here, we investigate this question by focusing on a single lineage of polyomaviruses that infect both humans and their closest relatives, the African great apes. We show that these viruses exhibit considerable host specificity and that their evolution largely mirrors that of their hosts, suggesting that codivergence with their hosts played a major role in their diversification. Our results provide statistical evidence in favor of an association of polyomaviruses and their hosts over millions of years. PMID:27440885

  3. Report of the first international workshop on human chromosome 14 mapping 1993

    SciTech Connect

    Cox, D.W.

    1995-06-01

    The first International Workshop on Human Chromosome 14 mapping was held at Novotel in Toronto, Canada on June 9-12, 1993. There were 23 participants from nine countries. The goals of the workshop were to compile physical maps and a consensus linkage map, to consolidate available data on disease loci, to catalogue and facilitate distribution of resources and to encourage new collaborations and data sharing.

  4. Identification and Characterization of Novel Rat Polyomavirus 2 in a Colony of X-SCID Rats by P-PIT assay

    PubMed Central

    Rigatti, Lora H.; Toptan, Tuna; Newsome, Joseph T.

    2016-01-01

    ABSTRACT Polyomaviruses (PyVs) are known to infect a wide range of vertebrates and invertebrates and are associated with a broad spectrum of diseases, including cancers, particularly in immune-suppressed hosts. A novel polyomavirus, designated rat polyomavirus 2 (RatPyV2), was identified from a breeding colony of rats having X-linked severe combined immunodeficiency. Using a human panpolyomavirus immunohistochemistry test (P-PIT), RatPyV2 was initially detected in the parotid salivary gland of a colony member. Rolling circle amplification using DNA from harderian and parotid glands identified a novel 5.1-kb polyomavirus genome closely related to human Washington University (WU) and Karolinska Institute (KI) and vole polyomaviruses but notably divergent from Rattus norvegicus PyV1 (RnorPyV1; also designated RatPyV1). Further screening showed RatPyV2 inclusion body infection in the lung epithelium and variably in other respiratory, reproductive, and glandular tissues of 12/12 (100%) rats. IMPORTANCE Although P-PIT was developed to detect diseases associated with known human polyomaviruses, the identification of a new polyomavirus in rats suggests that it may have utility as a broad-based screen for new, as well as known polyomaviruses. Our findings suggest that RatPyV2 may be a commensal infection of laboratory rats that can lead to disseminated disease in T cell immune-deficient rats. Infection of the X-SCID rats with RatPyV2 and Pneumocystis carinii is a potential model for coinfection pathogenesis and treatment options during transplant preclinical studies. PMID:28028546

  5. The Merkel Cell Polyomavirus Minor Capsid Protein

    PubMed Central

    Schowalter, Rachel M.; Buck, Christopher B.

    2013-01-01

    The surface of polyomavirus virions is composed of pentameric knobs of the major capsid protein, VP1. In previously studied polyomavirus species, such as SV40, two interior capsid proteins, VP2 and VP3, emerge from the virion to play important roles during the infectious entry process. Translation of the VP3 protein initiates at a highly conserved Met-Ala-Leu motif within the VP2 open reading frame. Phylogenetic analyses indicate that Merkel cell polyomavirus (MCV or MCPyV) is a member of a divergent clade of polyomaviruses that lack the conserved VP3 N-terminal motif. Consistent with this observation, we show that VP3 is not detectable in MCV-infected cells, VP3 is not found in native MCV virions, and mutation of possible alternative VP3-initiating methionine codons did not significantly affect MCV infectivity in culture. In contrast, VP2 knockout resulted in a >100-fold decrease in native MCV infectivity, despite normal virion assembly, viral DNA packaging, and cell attachment. Although pseudovirus-based experiments confirmed that VP2 plays an essential role for infection of some cell lines, other cell lines were readily transduced by pseudovirions lacking VP2. In cell lines where VP2 was needed for efficient infectious entry, the presence of a conserved myristoyl modification on the N-terminus of VP2 was important for its function. The results show that a single minor capsid protein, VP2, facilitates a post-attachment stage of MCV infectious entry into some, but not all, cell types. PMID:23990782

  6. Human Genome Diversity Project. Summary of planning workshop 3(B): Ethical and human-rights implications

    SciTech Connect

    1993-12-31

    The third planning workshop of the Human Genome Diversity Project was held on the campus of the US National Institutes of Health in Bethesda, Maryland, from February 16 through February 18, 1993. The second day of the workshop was devoted to an exploration of the ethical and human-rights implications of the Project. This open meeting centered on three roundtables, involving 12 invited participants, and the resulting discussions among all those present. Attendees and their affiliations are listed in the attached Appendix A. The discussion was guided by a schedule and list of possible issues, distributed to all present and attached as Appendix B. This is a relatively complete, and thus lengthy, summary of the comments at the meeting. The beginning of the summary sets out as conclusions some issues on which there appeared to be widespread agreement, but those conclusions are not intended to serve as a set of detailed recommendations. The meeting organizer is distributing his recommendations in a separate memorandum; recommendations from others who attended the meeting are welcome and will be distributed by the meeting organizer to the participants and to the Project committee.

  7. Achievable Human Exploration of Mars: Highlights from The Fourth Community Workshop (AM IV)

    NASA Technical Reports Server (NTRS)

    Thronson, Harley; Cassady, Joseph

    2017-01-01

    About a half decade ago, several professionals working mainly in industry on scenarios for initial human exploration of Mars together recognized that, under generally similar assumptions, there was a fair degree of similarity among these scenarios. Moreover, opportunities should be sought for greater community input into NASA's own scenario-building for the future of human space flight. A series of focused community workshops were considered to be effective to assess these scenarios and involve more directly the science community, including planetary protection, with industry. Four workshops to date each involve about sixty professional scientists, engineers, technologists, and strategists from NASA, academia, aerospace corporations, the National Academies, consulting organizations, and potential international partners.

  8. Polymerase chain reaction assay for avian polyomavirus.

    PubMed

    Phalen, D N; Wilson, V G; Graham, D L

    1991-05-01

    A polymerase chain reaction assay was developed for detection of budgerigar fledgling disease virus (BFDV). The assay used a single set of primers complementary to sequences located in the putative coding region for the BFDV VP1 gene. The observed amplification product had the expected size of 550 bp and was confirmed to derive from BFDV DNA by its restriction digestion pattern. This assay was specific for BFDV and highly sensitive, being able to detect as few as 20 copies of the virus. By using the polymerase chain reaction, BFDV was detected in adult, nestling, and embryo budgerigar (Melopsitticus undulatus) tissue DNAs and in sera from adult and nestling budgerigars. These results suggest the possibility of persistent infections in adult birds and lend further support to previously described evidence of possible in ovo transmission. BFDV was also detected in chicken embryo fibroblast cell cultures and chicken eggs inoculated with the virus. A 550-bp product with identical restriction enzyme sites was amplified from a suspected polyomavirus isolated from a peach-faced lovebird (Agapornis pesonata) and from tissue DNA from a Hahn's macaw (Ara nobilis) and a sun conure (Aratinga solstitialis) with histological lesions suggestive of polyomavirus infection. These fragments also hybridized with a BFDV-derived probe, proving that they were derived from a polyomavirus very similar, if not identical, to BFDV.

  9. Polymerase chain reaction assay for avian polyomavirus.

    PubMed Central

    Phalen, D N; Wilson, V G; Graham, D L

    1991-01-01

    A polymerase chain reaction assay was developed for detection of budgerigar fledgling disease virus (BFDV). The assay used a single set of primers complementary to sequences located in the putative coding region for the BFDV VP1 gene. The observed amplification product had the expected size of 550 bp and was confirmed to derive from BFDV DNA by its restriction digestion pattern. This assay was specific for BFDV and highly sensitive, being able to detect as few as 20 copies of the virus. By using the polymerase chain reaction, BFDV was detected in adult, nestling, and embryo budgerigar (Melopsitticus undulatus) tissue DNAs and in sera from adult and nestling budgerigars. These results suggest the possibility of persistent infections in adult birds and lend further support to previously described evidence of possible in ovo transmission. BFDV was also detected in chicken embryo fibroblast cell cultures and chicken eggs inoculated with the virus. A 550-bp product with identical restriction enzyme sites was amplified from a suspected polyomavirus isolated from a peach-faced lovebird (Agapornis pesonata) and from tissue DNA from a Hahn's macaw (Ara nobilis) and a sun conure (Aratinga solstitialis) with histological lesions suggestive of polyomavirus infection. These fragments also hybridized with a BFDV-derived probe, proving that they were derived from a polyomavirus very similar, if not identical, to BFDV. Images PMID:1647403

  10. Human genetics for non-scientists: Practical workshops for policy makers and opinion leaders

    SciTech Connect

    1995-12-31

    These workshops form part of a series of workshops that the Banbury and the DNA Learning Centers of Cold Spring Harbor Laboratory have held for a number of years, introducing genetics, and the ways in which scientific research is done, to non-scientists. The purpose of the workshops as stated in the grant application was: {open_quotes}Our objective is to foster a better understanding of the societal impact of human genome research by providing basic information on genetics to non-scientists whose professions or special interests interface with genetic technology.... Participants will be chosen for their interest in human genetics and for their roles as opinion leaders in their own communities. Primary care physicians are of particular interest to us for this series of workshops.{close_quotes} Two workshops were held under this grant. The first was held in 21-24 April, 1994 and attended by 20 participants, and the second was held 16-19 November, 1995, and attended by 16 participants. In each case, there was a combination of concept lectures on the foundations of human molecular genetics; lectures by invited specialists; and laboratory experiments to introduce non-scientists to the techniques used in molecular genetics.

  11. Oncogenic Papillomavirus and Polyomavirus in Water Environments: Is There a Potential for Waterborne Transmission?

    PubMed

    Fratini, M; Di Bonito, P; La Rosa, G

    2014-03-01

    Waterborne exposure to human viruses through contact with sewage-contaminated water environments can result in infections associated with a wide range of illnesses. Gastrointestinal symptoms are the most commonly encountered manifestations of waterborne viral illness. Respiratory diseases, neurological diseases and paralysis can also occur. Whether viral infections resulting in health outcomes like cancer might also be transmitted by the waterborne route is unknown. Recently, viruses belonging to two oncogenic groups-Human Papillomaviruses (HPVs) and Human Polyomaviruses (HPyVs)-have been detected in urban sewages worldwide. The latter have also been identified in other water environments. HPVs are epitheliotropic viruses responsible for several diseases of skin and mucosae, from common warts to squamous intraepithelial lesions that can either heal or progress to invasive carcinoma of the cervix, vulva, vagina, penis, anus or oropharynx. Human PyVs infect different tissues and organs, causing infections that are usually subclinical in immunocompetent individuals but can be serious in immunocompromised hosts. These pathogens belong to a family of DNA tumour viruses. Merkel cell polyomavirus, a HPyV identified in recent years, has attracted much attention due to its link with a rare and aggressive form of human cancer. Merkel cell carcinoma, the incidence of which has tripled over the past two decades. JC polyomavirus and BK polyomavirus are also potentially oncogenic. The observed abundance and wide dissemination of HPVs and HPyVs in water environments strongly suggest the need to shed light on the fate of these viruses in water environments and to elucidate their potential for waterborne transmission. Such information is essential for the improvement of wastewater management programs in terms of both sewage treatment and water quality surveillance.

  12. Achieving And Sustaining Human Exploration of Mars The Fourth Community Workshop (AM IV)

    NASA Technical Reports Server (NTRS)

    Thronson, Harley; Beaty, David; Carberry, Chris; Drake, Bret; Hays, Lindsay

    2017-01-01

    About a half decade ago, several professionals working mainly in industry on scenarios for initial human exploration of Mars exploration together recognized that, under generally similar assumptions, there was a fair degree of similarity among these scenarios. Moreover, opportunities should be sought for greater community input into NASAs own scenario-building for the future of human space flight. A series of focused community workshops were considered to be effective to critically assess the increasingly sophisticated scenarios. Explore Mars, Inc. the American Astronautical Society agreed to support them. Four workshops to date each involve about sixty professional scientists, engineers, technologists, and strategists from NASA, academia, aerospace corporations, the National Academies, consulting organizations, and potential international partners. Each workshop produced a series of presentations and reports briefed to NASA leadership and other stakeholders.

  13. How Polyomaviruses Exploit the ERAD Machinery to Cause Infection

    PubMed Central

    Dupzyk, Allison; Tsai, Billy

    2016-01-01

    To infect cells, polyomavirus (PyV) traffics from the cell surface to the endoplasmic reticulum (ER) where it hijacks elements of the ER-associated degradation (ERAD) machinery to penetrate the ER membrane and reach the cytosol. From the cytosol, the virus transports to the nucleus, enabling transcription and replication of the viral genome that leads to lytic infection or cellular transformation. How PyV exploits the ERAD machinery to cross the ER membrane and access the cytosol, a decisive infection step, remains enigmatic. However, recent studies have slowly unraveled many aspects of this process. These emerging insights should advance our efforts to develop more effective therapies against PyV-induced human diseases. PMID:27589785

  14. Report on the COSPAR Workshop on Refining Planetary Protection Requirements for Human Missions

    NASA Astrophysics Data System (ADS)

    Spry, James Andrew; Rummel, John; Conley, Catharine; Race, Margaret; Kminek, Gerhard; Siegel, Bette

    2016-07-01

    A human mission to Mars has been the driving long-term goal for the development of the Global Exploration Roadmap by the International Space Exploration Coordination Group. Additionally, multiple national space agencies and commercial organizations have published similar plans and aspirations for human missions beyond LEO. The current COSPAR planetary protection "Guidelines for Human Missions to Mars" were developed in a series of workshops in the early 2000s and adopted into COSPAR policy at the Montreal Assembly in 2008. With changes and maturation in mission architecture concepts and hardware capabilities, the holding of a workshop provided an opportunity for timely review of these guidelines and their interpretation within current frameworks provided by ISECG and others. The COSPAR Workshop on Refining Planetary Protection Requirements for Human Missions was held in the US in spring 2016 to evaluate recent efforts and activities in the context of current COSPAR policy, as well as collect inputs from the various organizations considering crewed exploration missions to Mars and precursor robotic missions focused on surface material properties and environmental challenges. The workshop also considered potential updates to the COSPAR policy for human missions across a range of planetary destinations. This paper will report on those deliberations.

  15. Identification of a Novel Polyomavirus from Patients with Acute Respiratory Tract Infections

    PubMed Central

    Gaynor, Anne M; Nissen, Michael D; Whiley, David M; Mackay, Ian M; Lambert, Stephen B; Wu, Guang; Brennan, Daniel C; Storch, Gregory A; Sloots, Theo P; Wang, David

    2007-01-01

    We report the identification of a novel polyomavirus present in respiratory secretions from human patients with symptoms of acute respiratory tract infection. The virus was initially detected in a nasopharyngeal aspirate from a 3-year-old child from Australia diagnosed with pneumonia. A random library was generated from nucleic acids extracted from the nasopharyngeal aspirate and analyzed by high throughput DNA sequencing. Multiple DNA fragments were cloned that possessed limited homology to known polyomaviruses. We subsequently sequenced the entire virus genome of 5,229 bp, henceforth referred to as WU virus, and found it to have genomic features characteristic of the family Polyomaviridae. The genome was predicted to encode small T antigen, large T antigen, and three capsid proteins: VP1, VP2, and VP3. Phylogenetic analysis clearly revealed that the WU virus was divergent from all known polyomaviruses. Screening of 2,135 patients with acute respiratory tract infections in Brisbane, Queensland, Australia, and St. Louis, Missouri, United States, using WU virus–specific PCR primers resulted in the detection of 43 additional specimens that contained WU virus. The presence of multiple instances of the virus in two continents suggests that this virus is geographically widespread in the human population and raises the possibility that the WU virus may be a human pathogen. PMID:17480120

  16. One Health Approach to Identify Research Needs in Bovine and Human Babesioses: Workshop Report

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Babesia are emerging health threats to humans and animals in the United States. A collaborative effort of multiple disciplines to attain optimal health for people, animals and our environment, otherwise known as the One Health concept, was taken during a research workshop held in April 2009 to iden...

  17. Report of the Second International Workshop on Human Chromosome 5 Mapping

    SciTech Connect

    Westbrook, C.A.; Neuman, W.L.; McPherson, J.; Wasmuth, J.; Camper, S.; Plaetke, R.; Williamson, R.

    1993-12-31

    This report describes the accomplishments of the Second International Workshop on Human Chromosome 5 as was held May 11--13,1992 at the University of Chicago. Included in the report are abstract of individual presentations and a consensus map of the chromosome.

  18. Human Factors of Flight-deck Automation: NASA/Industry Workshop

    NASA Technical Reports Server (NTRS)

    Boehm-Davis, D. A.; Curry, R. E.; Wiener, E. L.; Harrison, R. L.

    1981-01-01

    The scope of automation, the benefits of automation, and automation-induced problems were discussed at a workshop held to determine whether those functions previously performed manually on the flight deck of commercial aircraft should always be automated in view of various human factors. Issues which require research for resolution were identified. The research questions developed are presented.

  19. Workshop on Mars 2001: Integrated Science in Preparation for Sample Return and Human Exploration

    NASA Technical Reports Server (NTRS)

    Marshall, John (Editor); Weitz, Cathy (Editor)

    1999-01-01

    The Workshop on Mars 2001: Integrated Science in Preparation for Sample Return and Human Exploration was held on October 2-4, 1999, at the Lunar and Planetary Institute in Houston, Texas. The workshop was sponsored by the Lunar and Planetary Institute, the Mars Program Office of the Jet Propulsion Laboratory, and the National Aeronautics and Space Administration. The three-day meeting was attended by 133 scientists whose purpose was to share results from recent missions, to share plans for the 2001 mission, and to come to an agreement on a landing site for this mission.

  20. Identification of the Neutralizing Epitopes of Merkel Cell Polyomavirus Major Capsid Protein within the BC and EF Surface Loops

    PubMed Central

    Fleury, Maxime J. J.; Nicol, Jérôme T. J.; Samimi, Mahtab; Arnold, Françoise; Cazal, Raphael; Ballaire, Raphaelle; Mercey, Olivier; Gonneville, Hélène; Combelas, Nicolas; Vautherot, Jean-Francois; Moreau, Thierry; Lorette, Gérard; Coursaget, Pierre; Touzé, Antoine

    2015-01-01

    Merkel cell polyomavirus (MCPyV) is the first polyomavirus clearly associated with a human cancer, i.e. the Merkel cell carcinoma (MCC). Polyomaviruses are small naked DNA viruses that induce a robust polyclonal antibody response against the major capsid protein (VP1). However, the polyomavirus VP1 capsid protein epitopes have not been identified to date. The aim of this study was to identify the neutralizing epitopes of the MCPyV capsid. For this goal, four VP1 mutants were generated by insertional mutagenesis in the BC, DE, EF and HI loops between amino acids 88-89, 150-151, 189-190, and 296-297, respectively. The reactivity of these mutants and wild-type VLPs was then investigated with anti-VP1 monoclonal antibodies and anti-MCPyV positive human sera. The findings together suggest that immunodominant conformational neutralizing epitopes are present at the surface of the MCPyV VLPs and are clustered within BC and EF loops. PMID:25812141

  1. Affordable Exploration of Mars: Recommendations from a Community Workshop on Sustainable Initial Human Missions

    NASA Technical Reports Server (NTRS)

    Thronson, Harley; Carberry, Chris; Cassady, R. J.; Cooke, Doug; Hopkins, Joshua; Perino, Maria A.; Kirkpatrick, Jim; Raftery, Michael; Westenberg, Artemis; Zucker, Richard

    2013-01-01

    There is a growing consensus that within two decades initial human missions to Mars are affordable under plausible budget assumptions and with sustained international participation. In response to this idea, a distinguished group of experts from the Mars exploration stakeholder communities attended the "Affording Mars" workshop at George Washington University in December, 2013. Participants reviewed and discussed scenarios for affordable and sustainable human and robotic exploration of Mars, the role of the International Space Station over the coming decade as the essential early step toward humans to Mars, possible "bridge" missions in the 2020s, key capabilities required for affordable initial missions, international partnerships, and a usable definition of affordability and sustainability. We report here the findings, observations, and recommendations that were agreed to at that workshop.

  2. Detection and genome characterization of bovine polyomaviruses in beef muscle and ground beef samples from Germany.

    PubMed

    Gräfe, Donina; Ehlers, Bernhard; Mäde, Dietrich; Ellerbroek, Lüppo; Seidler, Tassilo; Johne, Reimar

    2017-01-16

    Polyomaviruses are small, non-enveloped, circular double-stranded DNA viruses. Some polyomaviruses can induce tumors and cancer under certain circumstances. The bovine polyomaviruses (BPyV) 1-3 have been only scarcely analyzed so far. It was hypothesized that the consumption of beef meat containing polyomaviruses could contribute to the development of cancer in humans. In order to assess the distribution of the BPyV genome in meat from Germany, 101 beef muscle samples and 10 ground beef samples were analyzed here. A specific sample preparation method combined with or without rolling circle amplification (RCA), and BPyV-specific PCRs were developed and applied. BPyV-1 DNA was detected in 1/101 (1%) samples from beef meat and in 2/10 (20%) ground beef samples. BPyV-2 DNA was detected in 3/10 (30%) ground beef samples, whereas BPyV-3 was not detected in the samples. Application of RCA did not increase the detection rate in ground beef samples. Sequence analysis of the PCR products indicated the presence of BPyV-1, BPyV-2a and BPyV-2b. The whole genome of a BPyV-1 strain from ground beef meat showed 97.8% sequence identity to the BPyV-1 reference strain and that of a BPyV-2a strain from ground beef meet showed 99.9% sequence identity to strain 2aS11. It can be concluded that BPyV genomes can be frequently detected in ground beef samples, although higher sample numbers should be investigated in future to confirm this finding. Further studies should focus on the infectivity, tumorigenicity and heat resistance of the contained viruses in order to assess the risk of cancer induction through consumption of BPyVs present in beef products.

  3. Role of Cell-Type-Specific Endoplasmic Reticulum-Associated Degradation in Polyomavirus Trafficking

    PubMed Central

    Bennett, Shauna M.; Jiang, Mengxi

    2013-01-01

    BK polyomavirus (BKPyV) is a widespread human pathogen that establishes a lifelong persistent infection and can cause severe disease in immunosuppressed patients. BKPyV is a nonenveloped DNA virus that must traffic through the endoplasmic reticulum (ER) for productive infection to occur; however, it is unknown how BKPyV exits the ER before nuclear entry. In this study, we elucidated the role of the ER-associated degradation (ERAD) pathway during BKPyV intracellular trafficking in renal proximal tubule epithelial (RPTE) cells, a natural host cell. Using proteasome and ERAD inhibitors, we showed that ERAD is required for productive entry. Altered trafficking and accumulation of uncoated viral intermediates were detected by fluorescence in situ hybridization and indirect immunofluorescence in the presence of an inhibitor. Additionally, we detected a change in localization of partially uncoated virus within the ER during proteasome inhibition, from a BiP-rich area to a calnexin-rich subregion, indicating that BKPyV accumulated in an ER subcompartment. Furthermore, inhibiting ERAD did not prevent entry of capsid protein VP1 into the cytosol from the ER. By comparing the cytosolic entry of the related polyomavirus simian virus 40 (SV40), we found that dependence on the ERAD pathway for cytosolic entry varied between the polyomaviruses and between different cell types, namely, immortalized CV-1 cells and primary RPTE cells. PMID:23740996

  4. Glucocorticoids facilitate the stable transformation of embryonal rat fibroblasts by a polyomavirus large tumor antigen-deficient mutant.

    PubMed Central

    Martens, I; Nilsson, M; Magnusson, G; Linder, S

    1988-01-01

    The addition of glucocorticoids to the growth medium could substitute for the expression of the polyomavirus large tumor antigen in the transformation of rat fibroblasts in vitro. After transfection with a large tumor antigen-deficient mutant of polyomavirus, pbc1051, high-frequency permanent transformation was observed, if the cells were grown in medium containing dexamethasone. Growth of pbc1051-transfected rat fibroblasts was strictly dependent on the presence of glucocorticoids during the initial phase of transformation. In the second phase, the growth of pbc1051-transfected cells was stimulated by dexamethasone, but the hormone was not essential for growth. After approximately 10 weeks in culture, pbc1051-transfected cells had progressed to hormone independent growth. Rat embryo cells transfected with wild-type polyomavirus DNA had the second phase in which growth was stimulated by glucocorticoid, and after this phase growth was steroid independent. Addition of glucocorticoids to rat fibroblasts transfected with a plasmid encoding only the middle-sized tumor antigen resulted in only a weak stimulation of growth. In contrast, embryo cells transfected with a plasmid containing the human homologue of the cellular T24 Ha-ras gene linked to murine sarcoma virus and simian virus 40 enhancers could be efficiently established as cell lines in medium supplemented with glucocorticoids. The data suggest that, in the transformation of primary rodent cells by polyomavirus, the activity of large tumor antigen can be substituted for by stimulating normal cellular functions with dexamethasone. Images PMID:2840668

  5. Workshop Summaries

    ERIC Educational Resources Information Center

    Dandes, Herbert

    1970-01-01

    Workshop titles are: (1) "Authenticity in Communication ; (2) "Inter Cultural Communication ; (3) "Enticements to Eupsychia ; (4) "Psychoneurosensory Problems in Education ; (5) "Why Education for Family Life and Human Sexuality ; and (6) "Communication in the Hazards of Drug Abuse . (EK)

  6. Human-Centered Technology for Maintainability: Workshop Proceedings

    DTIC Science & Technology

    1991-06-01

    designer with several specific functions for evaluating work space and task configurations. The functions or design tasks are selected based on the...objective function , Schultz et al. (1981; 1982) found that such techniques produced good agreement between computed muscle tcnsion and measured ...constituent :ielcds such as computer-graphics, human modeling, arithropometric and related measurement pronlerns, v.orxstation concepts, and human

  7. Human Dimensions in Future Battle Command Systems: A Workshop Report

    DTIC Science & Technology

    2008-04-01

    on technology pedagogy while emphasizing professional self- and group- developments and experiential training. The training and education courses...technology with the human in-the-loop and partly operational in the level of autonomy with no human involvement. In both cases, information in BCS... pedagogy while emphasizing professional self- and group- developments and experiential training. The training and education courses should be

  8. Evolution of the BK polyomavirus: epidemiological, anthropological and clinical implications.

    PubMed

    Yogo, Yoshiaki; Sugimoto, Chie; Zhong, Shan; Homma, Yukio

    2009-07-01

    BK polyomavirus (BKV) is essentially ubiquitous in all human populations worldwide. Asymptomatic infection with this virus occurs during early childhood, leading to life-long persistence in the kidney. BKV has four subtypes that can be identified using serological and genotyping methods. The evolutionary aspects of BKV have remained poorly understood due to the limited availability of BKV genomes, since urinary excretion of BKV DNA is detected primarily in immunocompromised individuals. However, we have found that BKV DNA sequences can often be amplified from non-immunocompromised elderly individuals, using a highly sensitive polymerase chain reaction (PCR) with highly concentrated urinary DNA as the source of viral DNA. Using this approach, we have PCR-amplified and sequenced a large number of partial and complete BKV genomes from various human populations worldwide and conducted a series of evolutionary studies using these sequences. We have shown that subtypes I and IV evolved into four and six subgroups, respectively, with each having a close relationship with a particular human population. In addition, we have provided evidence supporting the hypothesis that BKV strains with the archetypal transcriptional control region (TCR) circulate in the human population. In this review, we describe these findings and discuss their epidemiological, anthropological and clinical implications.

  9. Expression and purification of recombinant polyomavirus VP2 protein and its interactions with polyomavirus proteins

    NASA Technical Reports Server (NTRS)

    Cai, X.; Chang, D.; Rottinghaus, S.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1994-01-01

    Recombinant polyomavirus VP2 protein was expressed in Escherichia coli (RK1448), using the recombinant expression system pFPYV2. Recombinant VP2 was purified to near homogeneity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electroelution, and Extracti-Gel chromatography. Polyclonal serum to this protein which reacted specifically with recombinant VP2 as well as polyomavirus virion VP2 and VP3 on Western blots (immunoblots) was produced. Purified VP2 was used to establish an in vitro protein-protein interaction assay with polyomavirus structural proteins and purified recombinant VP1. Recombinant VP2 interacted with recombinant VP1, virion VP1, and the four virion histones. Recombinant VP1 coimmunoprecipitated with recombinant VP2 or truncated VP2 (delta C12VP2), which lacked the carboxy-terminal 12 amino acids. These experiments confirmed the interaction between VP1 and VP2 and revealed that the carboxyterminal 12 amino acids of VP2 and VP3 were not necessary for formation of this interaction. In vivo VP1-VP2 interaction study accomplished by cotransfection of COS-7 cells with VP2 and truncated VP1 (delta N11VP1) lacking the nuclear localization signal demonstrated that VP2 was capable of translocating delta N11VP1 into the nucleus. These studies suggest that complexes of VP1 and VP2 may be formed in the cytoplasm and cotransported to the nucleus for virion assembly to occur.

  10. Cooperation between the polyomavirus Middle-T-antigen gene and the human c-myc oncogene in a rat thyroid epithelial differentiated cell line: Model of in vitro progression

    SciTech Connect

    Berlingieri, M.T.; Portella, G.; Grieco, M.; Santoro, M.; Fusco, A.

    1988-05-01

    Two rat thyroid epithelial differentiated cell lines, PC CI 3 and PC myc, were infected with the polyoma murine leukemia virus (PyMLV) carrying the Middle-T-antigen gene of polyomavirus. After infection, both cell lines acquired the typical markers of neoplastic transformation; however, the PC myc cells showed a greater malignant phenotype. Furthermore, the thyroid differentiated functions were completely suppressed in PC myc cells transformed by PyMLV, whereas they were, at least partially, retained in PC CI 3 cells transformed by PyMLV, and in particular, thyroglobulin synthesis and secretion were not affected at all. Since no differences in the expression of the middle-T-antigen gene were observed in the two PyMLV-transformed cell lines, the different properties shown by these two infected cell lines must be ascribed to the expression of the c-myc oncogene.

  11. Prioritizing Maintenance Human Factors Challenges and Solutions: Workshop Proceedings

    DTIC Science & Technology

    2011-08-01

    Bobby Reed ACE Mary Schooley ANM John Sims AAL Vickie A. Stahlberg AWP Michele Wallentine ANM Katherine Wilson NTSB Nadine Yeager ACE v CONTENTs...collective action items from the three presenters. Investigative Reports on Maintenance-Related Accidents. Dr. Katherine Wilson (Human Performance...failure to follow procedures; unqualified technician without task specific training; improper over- sight of work; and inadequate inspection. Dr. Wilson

  12. CD Nomenclature 2015: Human Leukocyte Differentiation Antigen Workshops as a Driving Force in Immunology.

    PubMed

    Engel, Pablo; Boumsell, Laurence; Balderas, Robert; Bensussan, Armand; Gattei, Valter; Horejsi, Vaclav; Jin, Bo-Quan; Malavasi, Fabio; Mortari, Frank; Schwartz-Albiez, Reinhard; Stockinger, Hannes; van Zelm, Menno C; Zola, Heddy; Clark, Georgina

    2015-11-15

    CD (cluster of differentiation) Ags are cell surface molecules expressed on leukocytes and other cells relevant for the immune system. CD nomenclature has been universally adopted by the scientific community and is officially approved by the International Union of Immunological Societies and sanctioned by the World Health Organization. It provides a unified designation system for mAbs, as well as for the cell surface molecules that they recognize. This nomenclature was established by the Human Leukocyte Differentiation Antigens Workshops. In addition to defining the CD nomenclature, these workshops have been instrumental in identifying and determining the expression and function of cell surface molecules. Over the past 30 y, the data generated by the 10 Human Leukocyte Differentiation Antigens Workshops have led to the characterization and formal designation of more than 400 molecules. CD molecules are commonly used as cell markers, allowing the identification and isolation of leukocyte populations, subsets, and differentiation stages. mAbs against these molecules have proven to be essential for biomedical research and diagnosis, as well as in biotechnology. More recently, they have been recognized as invaluable tools for the treatment of several malignancies and autoimmune diseases. In this article, we describe how the CD nomenclature was established, present the official updated list of CD molecules, and provide a rationale for their usefulness in the 21st century.

  13. Efficient propagation of archetype BK and JC polyomaviruses.

    PubMed

    Broekema, Nicole M; Imperiale, Michael J

    2012-01-20

    BKPyV and JCPyV are closely related, ubiquitous human pathogens that cause disease in immunocompromised patients. The DNA sequence of the regulatory regions distinguishes two forms of these viruses, designated archetype and rearranged. Although cell culture systems exist for rearranged BKPyV and JCPyV, currently there is no robust cell culture system to study the archetype viruses. Large T antigen (TAg) is a virally encoded protein required to initiate viral DNA synthesis. Because archetype virus produces undetectable levels of TAg, we hypothesized that TAg overexpression would stimulate archetype virus replication. Efficient propagation of the archetype forms of BKPyV and JCPyV was observed in 293TT cells, human embryonic kidney cells overexpressing SV40 TAg. Importantly, the archetypal structure of the regulatory region was maintained during viral growth. Significant replication was not observed for Merkel cell, KI, or WU polyomaviruses. 293TT cells provide a means of propagating archetype BKPyV and JCPyV for detailed study.

  14. The polyomavirus BK agnoprotein co-localizes with lipid droplets

    SciTech Connect

    Unterstab, Gunhild; Gosert, Rainer; Leuenberger, David; Lorentz, Pascal; Rinaldo, Christine H.; Hirsch, Hans H.

    2010-04-10

    Agnoprotein encoded by human polyomavirus BK (BKV) is a late cytoplasmic protein of 66 amino acids (aa) of unknown function. Immunofluorescence microscopy revealed a fine granular and a vesicular distribution in donut-like structures. Using BKV(Dunlop)-infected or agnoprotein-transfected cells, we investigated agnoprotein co-localization with subcellular structures. We found that agnoprotein co-localizes with lipid droplets (LD) in primary human renal tubular epithelial cells as well as in other cells supporting BKV replication in vitro (UTA, Vero cells). Using agnoprotein-enhanced green fluorescent protein (EGFP) fusion constructs, we demonstrate that agnoprotein aa 20-42 are required for targeting LD, whereas aa 1-20 or aa 42-66 were not. Agnoprotein aa 22-40 are predicted to form an amphipathic helix, and mutations A25D and F39E, disrupting its hydrophobic domain, prevented LD targeting. However, changing the phosphorylation site serine-11 to alanine or aspartic acid did not alter LD co-localization. Our findings provide new clues to unravel agnoprotein function.

  15. Planetary protection and humans missions to Mars: summary results from two workshops sponsored by NASA and NASA/ESA

    NASA Astrophysics Data System (ADS)

    Race, M. S.; Kminek, G.; Rummel, J. D.; Nasa; Nasa/Esa Workshop Participants

    Planetary Protection PP requirements will strongly influence mission and spacecraft designs for future human missions to Mars particularly those related to the operation of advanced life support systems ALS extravehicular activities EVA laboratory and in situ sampling operations and systems for environmental monitoring and control EMC In order to initiate communication understanding and working relations between the ALS EVA EMC and PP communities in both NASA and ESA two separate workshops were held to focus on mission-specific PP issues during future human missions to Mars The NASA Life Support and Habitation and Planetary Protection Workshop was held in Houston TX Center for Advanced Space Studies April 2005 and The Mars PP and Human Systems Research and Technology Joint NASA ESA Workshop was held at ESA ESTEC Noordwijk Netherlands May 2005 This poster presentation summarizes the findings of both workshops and their associated recommendations which are summarized as follows The NASA workshop developed a tentative conceptual approach consistent with current PP requirements to provide preliminary guidance in the assessment of EVA ALS EMC and other aspects of human missions The workshop report identified the need for development of a comprehensive classification and zoning system for Mars to minimize contamination and guide operations particularly in relation to COSPAR Special Region and protection of science and environmental conditions Critical research and technology

  16. Summary of a workshop on regulatory acceptance of (Q)SARs for human health and environmental endpoints.

    PubMed Central

    Jaworska, Joanna S; Comber, M; Auer, C; Van Leeuwen, C J

    2003-01-01

    The "Workshop on Regulatory Use of (Q)SARs for Human Health and Environmental Endpoints," organized by the European Chemical Industry Council and the International Council of Chemical Associations, gathered more than 60 human health and environmental experts from industry, academia, and regulatory agencies from around the world. They agreed, especially industry and regulatory authorities, that the workshop initiated great potential for the further development and use of predictive models, that is, quantitative structure-activity relationships [(Q)SARs], for chemicals management in a much broader scope than is currently the case. To increase confidence in (Q)SAR predictions and minimization of their misuse, the workshop aimed to develop proposals for guidance and acceptability criteria. The workshop also described the broad outline of a system that would apply that guidance and acceptability criteria to a (Q)SAR when used for chemical management purposes, including priority setting, risk assessment, and classification and labeling. PMID:12896859

  17. Diagnostic Assays for Polyomavirus JC and Progressive Multifocal Leukoencephalopathy

    PubMed Central

    White, Martyn K.; Sariyer, Ilker K.; Gordon, Jennifer; Delbue, Serena; Pietropaolo, Valeria; Berger, Joseph R.; Khalili, Kamel

    2016-01-01

    SUMMARY Progressive multifocal leukoencephalopathy (PML) is a devastating and often fatal demyelinating disease of the central nervous system (CNS) for which effective therapies are lacking. It is caused by the replication of polyomavirus JC (JCV) in the oligodendrocytes and astrocytes leading to their cytolytic death and loss of myelin from the subcortical white matter. While the virus is very common in human populations worldwide, the incidence of the disease is very low and confined almost exclusively to individuals with some form of immunological dysfunction. However, the number of people who constitute the at-risk population is growing larger and includes individuals with HIV-1/AIDS and patients receiving immunomodulatory therapies such as multiple sclerosis patients treated with natalizumab. Further adding to the public health significance of this disease are the difficulties encountered in the diagnosis of PML and the lack of useful biomarkers for PML progression. In this review, we examine the diagnostic assays that are available for different aspects of the JCV life cycle, their usefulness and drawbacks, and the prospects for improvements. PMID:26663440

  18. Immunohistochemical detection of KI polyomavirus in lung and spleen.

    PubMed

    Siebrasse, Erica A; Nguyen, Nang L; Smith, Colin; Simmonds, Peter; Wang, David

    2014-11-01

    Little is known about the tissue tropism of KI polyomavirus (KIPyV), and there are no studies to date describing any specific cell types it infects. The limited knowledge of KIPyV tropism has hindered study of this virus and understanding of its potential pathogenesis in humans. We describe tissues from two immunocompromised patients that stained positive for KIPyV antigen using a newly developed immunohistochemical assay targeting the KIPyV VP1 (KVP1) capsid protein. In the first patient, a pediatric bone marrow transplant recipient, KVP1 was detected in lung tissue. Double immunohistochemical staining demonstrated that approximately 50% of the KVP1-positive cells were CD68-positive cells of the macrophage/monocyte lineage. In the second case, an HIV-positive patient, KVP1 was detected in spleen and lung tissues. These results provide the first identification of a specific cell type in which KVP1 can be detected and expand our understanding of basic properties and in vivo tropism of KIPyV.

  19. Small-molecule inhibitors of JC polyomavirus infection

    PubMed Central

    Yatawara, Achani; Gaidos, Gabriel; Rupasinghe, Chamila N.; O’Hara, Bethany A.; Pellegrini, Maria; Atwood, Walter J.; Mierke, Dale F.

    2015-01-01

    The JC polyomavirus (JCPyV) infects approximately 50% of the human population. In healthy individuals the infection remains dormant and asymptomatic, but in immuno-suppressed patients it can cause progressive multifocal leukoencephalopathy (PML), a potentially fatal demyelinating disease. Currently, there are no drugs against JCPyV infection, nor for the treatment of PML. Here, we report the development of small molecule inhibitors of JCPyV that target the initial interaction between the virus and host cell and thereby block viral entry. Utilizing a combination of computational and NMR-based screening techniques, we target the LSTc tetrasaccharide binding site within the VP1 pentameric coat protein of JCPyV. Four of the compounds from the screen effectively block viral infection in our in vitro assays using SVG-A cells. For the most potent compound, we used saturation transfer difference NMR to determine the mode of binding to purified pentamers of JCPyV VP1. Collectively these results demonstrate the viability of this class of compounds for eventual development of JCPyV-antiviral therapeutics. PMID:25522925

  20. Probiotics in human milk and probiotic supplementation in infant nutrition: a workshop report.

    PubMed

    Bergmann, Henrike; Rodríguez, Juan Miguel; Salminen, Seppo; Szajewska, Hania

    2014-10-14

    Probiotics in human milk are a very recent field of research, as the existence of the human milk microbiome was discovered only about a decade ago. Current research is focusing on bacterial diversity and the influence of the maternal environment as well as the mode of delivery on human milk microbiota, the pathways of bacterial transfer to milk ducts, possible benefits of specific bacterial strains for the treatment of mastitis in mothers, and disease prevention in children. Recent advances in the assessment of early host-microbe interactions suggest that early colonisation may have an impact on later health. This review article summarises a scientific workshop on probiotics in human milk and their implications for infant health as well as future perspectives for infant feeding.

  1. The trichodysplasia spinulosa-associated polyomavirus: virological background and clinical implications.

    PubMed

    Kazem, Siamaque; van der Meijden, Els; Feltkamp, Mariet C W

    2013-08-01

    Trichodysplasia spinulosa-associated polyomavirus (TSPyV) is a new species of the family Polyomaviridae that was discovered in 2010. TSPyV infects humans and is associated with the development of a rare disease called trichodysplasia spinulosa. Trichodysplasia spinulosa is a skin disease of severely immunocompromised hosts characterized by follicular distention and keratotic spine formation especially on the face. Electron microscopy, immunohistochemistry, and viral load measurements indicate an etiological role of active TSPyV infection in the development of this disease. This review will address virological and pathogenic properties of TSPyV, as well as epidemiologic, diagnostic, and therapeutic aspects of TSPyV infection.

  2. The Science and Issues of Human DNA Polymorphisms: A Training Workshop for High School Biology Teachers

    SciTech Connect

    Micklos, David A.

    2006-10-30

    This project achieved its goal of implementing a nationwide training program to introduce high school biology teachers to the key uses and societal implications of human DNA polymorphisms. The 2.5-day workshop introduced high school biology faculty to a laboratory-based unit on human DNA polymorphisms â which provides a uniquely personal perspective on the science and Ethical, Legal and Social Implications (ELSI) of the Human Genome Project. As proposed, 12 workshops were conducted at venues across the United States. The workshops were attended by 256 high school faculty, exceeding proposed attendance of 240 by 7%. Each workshop mixed theoretical, laboratory, and computer work with practical and ethical implications. Program participants learned simplified lab techniques for amplifying three types of chromosomal polymorphisms: an Alu insertion (PV92), a VNTR (pMCT118/D1S80), and single nucleotide polymorphisms (SNPs) in the mitochondrial control region. These polymorphisms illustrate the use of DNA variations in disease diagnosis, forensic biology, and identity testing - and provide a starting point for discussing the uses and potential abuses of genetic technology. Participants also learned how to use their Alu and mitochondrial data as an entrée to human population genetics and evolution. Our work to simplify lab techniques for amplifying human DNA polymorphisms in educational settings culminated with the release in 1998 of three Advanced Technology (AT) PCR kits by Carolina Biological Supply Company, the nationâÂÂs oldest educational science supplier. The kits use a simple 30-minute method to isolate template DNA from hair sheaths or buccal cells and streamlined PCR chemistry based on Pharmacia Ready-To-Go Beads, which incorporate Taq polymerase, deoxynucleotide triphosphates, and buffer in a freeze-dried pellet. These kits have greatly simplified teacher implementation of human PCR labs, and their use is growing at a rapid pace. Sales of human

  3. The Science and Issues of Human DNA Polymoprhisms: A Training Workshop for High School Biology Teachers

    SciTech Connect

    David. A Micklos

    2006-10-30

    This project achieved its goal of implementing a nationwide training program to introduce high school biology teachers to the key uses and societal implications of human DNA polymorphisms. The 2.5-day workshop introduced high school biology faculty to a laboratory-based unit on human DNA polymorphisms – which provides a uniquely personal perspective on the science and Ethical, Legal and Social Implications (ELSI) of the Human Genome Project. As proposed, 12 workshops were conducted at venues across the United States. The workshops were attended by 256 high school faculty, exceeding proposed attendance of 240 by 7%. Each workshop mixed theoretical, laboratory, and computer work with practical and ethical implications. Program participants learned simplified lab techniques for amplifying three types of chromosomal polymorphisms: an Alu insertion (PV92), a VNTR (pMCT118/D1S80), and single nucleotide polymorphisms (SNPs) in the mitochondrial control region. These polymorphisms illustrate the use of DNA variations in disease diagnosis, forensic biology, and identity testing - and provide a starting point for discussing the uses and potential abuses of genetic technology. Participants also learned how to use their Alu and mitochondrial data as an entrée to human population genetics and evolution. Our work to simplify lab techniques for amplifying human DNA polymorphisms in educational settings culminated with the release in 1998 of three Advanced Technology (AT) PCR kits by Carolina Biological Supply Company, the nation’s oldest educational science supplier. The kits use a simple 30-minute method to isolate template DNA from hair sheaths or buccal cells and streamlined PCR chemistry based on Pharmacia Ready-To-Go Beads, which incorporate Taq polymerase, deoxynucleotide triphosphates, and buffer in a freeze-dried pellet. These kits have greatly simplified teacher implementation of human PCR labs, and their use is growing at a rapid pace. Sales of human polymorphism

  4. Qualitative and quantitative comparability of human and animal developmental neurotoxicants: a workshop summary.

    PubMed

    Rees, D C; Francis, E Z; Kimmel, C A

    1990-01-01

    A Workshop on the Qualitative and Quantitative Comparability of Human and Animal Developmental Neurotoxicity was held in Williamsburg, Va. on April 11-13, 1989. Based upon data presented at the Workshop, the degree of qualitative and quantitative comparability between data obtained from humans and experimental animals is reviewed for several developmental neurotoxicants (lead, agents of abuse, alcohol, PCBs, phenytoin, methylmercury, and ionizing radiation). Qualitative comparability was considered for the following functional categories: motor development and function, cognitive function, sensory function, motivation/arousal behavior, and social behavior. Quantitative comparability was assessed by comparing administered dose as well as measures of internal dose (e.g., blood levels) for selected agents. Comparability of qualitative changes between humans and rodents was most apparent when comparisons were made on the basis of general categories of behavioral function. These data support the use of animal models in assessing risk for developmental neurotoxicants and provide guidance on the types of functional end points that can be incorporated into a developmental neurotoxicity testing battery. Evidence of quantitative comparability was most apparent when an internal measure of dose (e.g., blood level) was used.

  5. New achievements in human cell toxicology: the 20th annual workshop on in vitro toxicology.

    PubMed

    Kolman, Ada

    2003-01-01

    The 20th Annual Workshop on In Vitro Toxicology (Oxford, UK, September 22-24, 2002) was convened as part of a European meeting entitled Human Cell Culture 2002. The meeting was arranged by the Scandinavian Society for Cell Toxicology (SSCT), the European Tissue Culture Society and the British Prostate Group. Two sessions, which are summarised in this report, were devoted to in vitro toxicology: Human Cell Toxicology and The SSCT Free Paper Session. Outstanding experts in the field of toxicology outlined contemporary approaches in toxicity testing in their lectures. Short oral presentations demonstrated a variety of in vitro model systems and methodologies, which can be useful for investigating human toxicity, as well as for studies on mechanisms of toxicity.

  6. Workshop on Critical Issues in Microgravity Fluids, Transport, and Reaction Processes in Advanced Human Support Technology

    NASA Technical Reports Server (NTRS)

    Chiaramonte, Francis P.; Joshi, Jitendra A.

    2004-01-01

    This workshop was designed to bring the experts from the Advanced Human Support Technologies communities together to identify the most pressing and fruitful areas of research where success hinges on collaborative research between the two communities. Thus an effort was made to bring together experts in both advanced human support technologies and microgravity fluids, transport and reaction processes. Expertise was drawn from academia, national laboratories, and the federal government. The intent was to bring about a thorough exchange of ideas and develop recommendations to address the significant open design and operation issues for human support systems that are affected by fluid physics, transport and reaction processes. This report provides a summary of key discussions, findings, and recommendations.

  7. Human factors issues in the use of artificial intelligence in air traffic control. October 1990 Workshop

    NASA Technical Reports Server (NTRS)

    Hockaday, Stephen; Kuhlenschmidt, Sharon (Editor)

    1991-01-01

    The objective of the workshop was to explore the role of human factors in facilitating the introduction of artificial intelligence (AI) to advanced air traffic control (ATC) automation concepts. AI is an umbrella term which is continually expanding to cover a variety of techniques where machines are performing actions taken based upon dynamic, external stimuli. AI methods can be implemented using more traditional programming languages such as LISP or PROLOG, or they can be implemented using state-of-the-art techniques such as object-oriented programming, neural nets (hardware or software), and knowledge based expert systems. As this technology advances and as increasingly powerful computing platforms become available, the use of AI to enhance ATC systems can be realized. Substantial efforts along these lines are already being undertaken at the FAA Technical Center, NASA Ames Research Center, academic institutions, industry, and elsewhere. Although it is clear that the technology is ripe for bringing computer automation to ATC systems, the proper scope and role of automation are not at all apparent. The major concern is how to combine human controllers with computer technology. A wide spectrum of options exists, ranging from using automation only to provide extra tools to augment decision making by human controllers to turning over moment-by-moment control to automated systems and using humans as supervisors and system managers. Across this spectrum, it is now obvious that the difficulties that occur when tying human and automated systems together must be resolved so that automation can be introduced safely and effectively. The focus of the workshop was to further explore the role of injecting AI into ATC systems and to identify the human factors that need to be considered for successful application of the technology to present and future ATC systems.

  8. Viral DNA Replication-Dependent DNA Damage Response Activation during BK Polyomavirus Infection

    PubMed Central

    Verhalen, Brandy; Justice, Joshua L.; Imperiale, Michael J.

    2015-01-01

    ABSTRACT BK polyomavirus (BKPyV) reactivation is associated with severe human disease in kidney and bone marrow transplant patients. The interplay between viral and host factors that regulates the productive infection process remains poorly understood. We have previously reported that the cellular DNA damage response (DDR) is activated upon lytic BKPyV infection and that its activation is required for optimal viral replication in primary kidney epithelial cells. In this report, we set out to determine what viral components are responsible for activating the two major phosphatidylinositol 3-kinase-like kinases (PI3KKs) involved in the DDR: ataxia telangiectasia mutated (ATM) kinase and ATM and Rad3-related (ATR) kinase. Using a combination of UV treatment, lentivirus transduction, and mutant virus infection experiments, our results demonstrate that neither the input virus nor the expression of large T antigen (TAg) alone is sufficient to trigger the activation of ATM or ATR in our primary culture model. Instead, our data suggest that the activation of both the ATM- and ATR-mediated DDR pathways is linked to viral DNA replication. Intriguingly, a TAg mutant virus that is unable to activate the DDR causes substantial host DNA damage. Our study provides insight into how DDRs are activated by polyomaviruses in primary cells with intact cell cycle checkpoints and how the activation might be linked to the maintenance of host genome stability. IMPORTANCE Polyomaviruses are opportunistic pathogens that are associated with several human diseases under immunosuppressed conditions. BK polyomavirus (BKPyV) affects mostly kidney and bone marrow transplant patients. The detailed replication mechanism of these viruses remains to be determined. We have previously reported that BKPyV activates the host DNA damage response (DDR), a response normally used by the host cell to combat genotoxic stress, to aid its own replication. In this study, we identified that the trigger for DDR

  9. Systems approach to understanding electromechanical activity in the human heart: a national heart, lung, and blood institute workshop summary.

    PubMed

    Rudy, Yoram; Ackerman, Michael J; Bers, Donald M; Clancy, Colleen E; Houser, Steven R; London, Barry; McCulloch, Andrew D; Przywara, Dennis A; Rasmusson, Randall L; Solaro, R John; Trayanova, Natalia A; Van Wagoner, David R; Varró, András; Weiss, James N; Lathrop, David A

    2008-09-09

    The National Heart, Lung, and Blood Institute (NHLBI) convened a workshop of cardiologists, cardiac electrophysiologists, cell biophysicists, and computational modelers on August 20 and 21, 2007, in Washington, DC, to advise the NHLBI on new research directions needed to develop integrative approaches to elucidate human cardiac function. The workshop strove to identify limitations in the use of data from nonhuman animal species for elucidation of human electromechanical function/activity and to identify what specific information on ion channel kinetics, calcium handling, and dynamic changes in the intracellular/extracellular milieu is needed from human cardiac tissues to develop more robust computational models of human cardiac electromechanical activity. This article summarizes the workshop discussions and recommendations on the following topics: (1) limitations of animal models and differences from human electrophysiology, (2) modeling ion channel structure/function in the context of whole-cell electrophysiology, (3) excitation-contraction coupling and regulatory pathways, (4) whole-heart simulations of human electromechanical activity, and (5) what human data are currently needed and how to obtain them. The recommendations can be found on the NHLBI Web site at http://www.nhlbi.nih.gov/meetings/workshops/electro.htm.

  10. Discovery of STL polyomavirus, a polyomavirus of ancestral recombinant origin that encodes a unique T antigen by alternative splicing.

    PubMed

    Lim, Efrem S; Reyes, Alejandro; Antonio, Martin; Saha, Debasish; Ikumapayi, Usman N; Adeyemi, Mitchell; Stine, O Colin; Skelton, Rebecca; Brennan, Daniel C; Mkakosya, Rajhab S; Manary, Mark J; Gordon, Jeffrey I; Wang, David

    2013-02-20

    The family Polyomaviridae is comprised of circular double-stranded DNA viruses, several of which are associated with diseases, including cancer, in immunocompromised patients. Here we describe a novel polyomavirus recovered from the fecal microbiota of a child in Malawi, provisionally named STL polyomavirus (STLPyV). We detected STLPyV in clinical stool specimens from USA and The Gambia at up to 1% frequency. Complete genome comparisons of two STLPyV strains demonstrated 5.2% nucleotide divergence. Alternative splicing of the STLPyV early region yielded a unique form of T antigen, which we named 229T, in addition to the expected large and small T antigens. STLPyV has a mosaic genome and shares an ancestral recombinant origin with MWPyV. The discovery of STLPyV highlights a novel alternative splicing strategy and advances our understanding of the complex evolutionary history of polyomaviruses.

  11. Structures of B-lymphotropic polyomavirus VP1 in complex with oligosaccharide ligands.

    PubMed

    Neu, Ursula; Khan, Zaigham Mahmood; Schuch, Benjamin; Palma, Angelina S; Liu, Yan; Pawlita, Michael; Feizi, Ten; Stehle, Thilo

    2013-10-01

    B-Lymphotropic Polyomavirus (LPyV) serves as a paradigm of virus receptor binding and tropism, and is the closest relative of the recently discovered Human Polyomavirus 9 (HPyV9). LPyV infection depends on sialic acid on host cells, but the molecular interactions underlying LPyV-receptor binding were unknown. We find by glycan array screening that LPyV specifically recognizes a linear carbohydrate motif that contains α2,3-linked sialic acid. High-resolution crystal structures of the LPyV capsid protein VP1 alone and in complex with the trisaccharide ligands 3'-sialyllactose and 3'-sialyl-N-acetyl-lactosamine (3SL and 3SLN, respectively) show essentially identical interactions. Most contacts are contributed by the sialic acid moiety, which is almost entirely buried in a narrow, preformed cleft at the outer surface of the capsid. The recessed nature of the binding site on VP1 and the nature of the observed glycan interactions differ from those of related polyomaviruses and most other sialic acid-binding viruses, which bind sialic acid in shallow, more exposed grooves. Despite their different modes for recognition, the sialic acid binding sites of LPyV and SV40 are half-conserved, hinting at an evolutionary strategy for diversification of binding sites. Our analysis provides a structural basis for the observed specificity of LPyV for linear glycan motifs terminating in α2,3-linked sialic acid, and links the different tropisms of known LPyV strains to the receptor binding site. It also serves as a useful template for understanding the ligand-binding properties and serological crossreactivity of HPyV9.

  12. 76 FR 60505 - Food Defense Workshop; Public Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-29

    ... HUMAN SERVICES Food and Drug Administration Food Defense Workshop; Public Workshop AGENCY: Food and Drug Administration, HHS. ACTION: Notice of public workshop. The Food and Drug Administration (FDA), Office of... M. Kerr Food & Agricultural Products Center (FAPC), is announcing a public workshop entitled...

  13. Workshop on human health impacts of halogenated biphenyls and related compounds.

    PubMed Central

    Kamrin, M A; Fischer, L J

    1991-01-01

    A workshop on the Human Health Impacts of Halogenated Biphenyls and Related Compounds was held to assess the state of current research on these chemicals and to make recommendations for future studies. Participants discussed results from laboratory animal experiments on PCBs, PBBs, dioxins, and dibenzofurans which demonstrate a common mode of toxicological action while also revealing large variations in toxicological potency both within and between these chemical families. These variations demonstrate the importance of congener-specific analyses in future studies of effects of exposure to these compounds. Results from epidemiological studies of environmentally exposed adult and pediatric populations from the U.S., Japan, and Taiwan and occupationally exposed cohorts from around the world were considered. It was concluded that available evidence did not demonstrate serious adverse effects such as cancer, in exposed adult cohorts but did provide indications of possible neurobehavioral effects in children exposed in utero. In addition, workshop participants described newly developed markers of exposure and techniques for assessing endocrinological, immunological, and neurological effects and suggested these be applied to epidemiological studies of the effects of polyhalogenated compounds. Other recommendations included identification of other cohorts and development of a large registry of exposed individuals; performance of detailed studies of reproductive function and outcomes in exposed populations; and follow up of neurobehavioral effects in offspring of exposed women. PMID:1674906

  14. Antarctic Exploration Parallels for Future Human Planetary Exploration: A Workshop Report

    NASA Technical Reports Server (NTRS)

    Hoffman, Stephen J. (Editor)

    2002-01-01

    Four Antarctic explorers were invited to a workshop at Johnson Space Center (JSC) to provide expert assessments of NASA's current understanding of future human exploration missions beyond low Earth orbit. These explorers had been on relatively sophisticated, extensive Antarctic expeditions with sparse or nonexistent support infrastructure in the period following World War II through the end of the International Geophysical Year. Their experience was similar to that predicted for early Mars or other planetary exploration missions. For example: one Antarctic a expedition lasted two years with only one planned resupply mission and contingency plans for no resupply missions should sea ice prevent a ship from reaching them; several traverses across Antarctica measured more than 1000 total miles, required several months to complete, and were made without maps (because they did not exist) and with only a few aerial photos of the route; and the crews of six to 15 were often international in composition. At JSC, the explorers were given tours of development, training, and scientific facilities, as well as documentation at operational scenarios for future planetary exploration. This report records their observations about these facilities and plans in answers to a series of questions provided to them before the workshop.

  15. Asymmetric assembly of Merkel cell polyomavirus large T-antigen origin binding domains at the viral origin.

    PubMed

    Harrison, Celia J; Meinke, Gretchen; Kwun, Hyun Jin; Rogalin, Henry; Phelan, Paul J; Bullock, Peter A; Chang, Yuan; Moore, Patrick S; Bohm, Andrew

    2011-06-17

    The double-stranded DNA polyomavirus Merkel cell polyomavirus (MCV) causes Merkel cell carcinoma, an aggressive but rare human skin cancer that most often affects immunosuppressed and elderly persons. As in other polyomaviruses, the large T-antigen of MCV recognizes the viral origin of replication by binding repeating G(A/G)GGC pentamers. The spacing, number, orientation, and necessity of repeats for viral replication differ, however, from other family members such as SV40 and murine polyomavirus. We report here the 2.9 Å crystal structure of the MCV large T-antigen origin binding domain (OBD) in complex with a DNA fragment from the MCV origin of replication. Consistent with replication data showing that three of the G(A/G)GGC-like binding sites near the center of the origin are required for replication, the crystal structure contains three copies of the OBD. This stoichiometry was verified using isothermal titration calorimetry. The affinity for G(A/G)GGC-containing double-stranded DNA was found to be ~740 nM, approximately 8-fold weaker than the equivalent domain in SV40 for the analogous region of the SV40 origin. The difference in affinity is partially attributable to DNA-binding residue Lys331 (Arg154 in SV40). In contrast to SV40, a small protein-protein interface is observed between MCV OBDs when bound to the central region of the origin. This protein-protein interface is reminiscent of that seen in bovine papilloma virus E1 protein. Mutational analysis indicates, however, that this interface contributes little to DNA binding energy.

  16. Asymmetric Assembly of Merkel Cell Polyomavirus Large T-Antigen Origin Binding Domains at the Viral Origin

    SciTech Connect

    C Harrison; G Meinke; H Kwun; H Rogalin; P Phelan; P Bullock; Y Chang; P Moore; A Bohm

    2011-12-31

    The double-stranded DNA polyomavirus Merkel cell polyomavirus (MCV) causes Merkel cell carcinoma, an aggressive but rare human skin cancer that most often affects immunosuppressed and elderly persons. As in other polyomaviruses, the large T-antigen of MCV recognizes the viral origin of replication by binding repeating G(A/G)GGC pentamers. The spacing, number, orientation, and necessity of repeats for viral replication differ, however, from other family members such as SV40 and murine polyomavirus. We report here the 2.9 {angstrom} crystal structure of the MCV large T-antigen origin binding domain (OBD) in complex with a DNA fragment from the MCV origin of replication. Consistent with replication data showing that three of the G(A/G)GGC-like binding sites near the center of the origin are required for replication, the crystal structure contains three copies of the OBD. This stoichiometry was verified using isothermal titration calorimetry. The affinity for G(A/G)GGC-containing double-stranded DNA was found to be {approx} 740 nM, approximately 8-fold weaker than the equivalent domain in SV40 for the analogous region of the SV40 origin. The difference in affinity is partially attributable to DNA-binding residue Lys331 (Arg154 in SV40). In contrast to SV40, a small protein-protein interface is observed between MCV OBDs when bound to the central region of the origin. This protein-protein interface is reminiscent of that seen in bovine papilloma virus E1 protein. Mutational analysis indicates, however, that this interface contributes little to DNA binding energy.

  17. Host range and cell cycle activation properties of polyomavirus large T-antigen mutants defective in pRB binding

    SciTech Connect

    Freund, R.; Bauer, P.H.; Benjamin, T.L.; Crissman, H.A.; Bradbury, E.M. |

    1994-11-01

    The authors have examined the growth properties of polyomavirus large T-antigen mutants that ar unable to bind pRB, the product of the retinoblastoma tumor suppressor gene. These mutants grow poorly on primary mouse cells yet grow well on NIH 3T3 and other established mouse cell lines. Preinfection of primary baby mouse kidney (BMK) epithelial cells with wild-type simian virus 40 renders these cells permissive to growth of pRB-binding polyomavirus mutants. Conversely, NIH 3T3 cells transfected by and expressing wild-type human pRB become nonpermissive. Primary fibroblasts for mouse embryos that carry a homozygous knockout of the RB gene are permissive, while those from normal littermates are nonpermissive. The host range of polyomavirus pRB-binding mutants is thus determined by expression or lack of expression of functional pRB by the host. These results demonstrate the importance of pRB binding by large T antigen for productive viral infection in primary cells. Failure of pRB-binding mutants to grow well in BMK cells correlates with their failure to induce progression from G{sub 0} or G{sub 1} through the S phase of the cell cycle. Time course studies show delayed synthesis and lower levels of accumulation of large T antigen, viral DNA, and VP1 in mutant compared with wild-type virus-infected BMK cells. These results support a model in which productive infection by polyomavirus in normal mouse cells is tightly coupled to the induction and progression of the cell cycle. 48 refs., 6 figs., 5 tabs.

  18. Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids.

    PubMed

    Ströh, Luisa J; Gee, Gretchen V; Blaum, Bärbel S; Dugan, Aisling S; Feltkamp, Mariet C W; Atwood, Walter J; Stehle, Thilo

    2015-08-01

    Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. Here we have used a structure-function approach to shed light on the first steps of TSPyV infection. We established by cell binding and pseudovirus infection studies that TSPyV interacts with sialic acids during attachment and/or entry. Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site.

  19. Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids

    PubMed Central

    Ströh, Luisa J.; Gee, Gretchen V.; Blaum, Bärbel S.; Dugan, Aisling S.; Feltkamp, Mariet C. W.; Atwood, Walter J.; Stehle, Thilo

    2015-01-01

    Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. Here we have used a structure-function approach to shed light on the first steps of TSPyV infection. We established by cell binding and pseudovirus infection studies that TSPyV interacts with sialic acids during attachment and/or entry. Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. PMID:26302170

  20. New Structural Insights into the Genome and Minor Capsid Proteins of BK Polyomavirus using Cryo-Electron Microscopy

    PubMed Central

    Hurdiss, Daniel L.; Morgan, Ethan L.; Thompson, Rebecca F.; Prescott, Emma L.; Panou, Margarita M.; Macdonald, Andrew; Ranson, Neil A.

    2016-01-01

    Summary BK polyomavirus is the causative agent of several diseases in transplant patients and the immunosuppressed. In order to better understand the structure and life cycle of BK, we produced infectious virions and VP1-only virus-like particles in cell culture, and determined their three-dimensional structures using cryo-electron microscopy (EM) and single-particle image processing. The resulting 7.6-Å resolution structure of BK and 9.1-Å resolution of the virus-like particles are the highest-resolution cryo-EM structures of any polyomavirus. These structures confirm that the architecture of the major structural protein components of these human polyomaviruses are similar to previous structures from other hosts, but give new insight into the location and role of the enigmatic minor structural proteins, VP2 and VP3. We also observe two shells of electron density, which we attribute to a structurally ordered part of the viral genome, and discrete contacts between this density and both VP1 and the minor capsid proteins. PMID:26996963

  1. The large tumor antigen: a "Swiss Army knife" protein possessing the functions required for the polyomavirus life cycle.

    PubMed

    Topalis, D; Andrei, G; Snoeck, R

    2013-02-01

    The SV40 large tumor antigen (L-Tag) is involved in the replication and cell transformation processes that take place during the polyomavirus life cycle. The ability of the L-Tag to interact with and to inactivate the tumor suppressor proteins p53 and pRb, makes this polyfunctional protein an interesting target in the search for compounds with antiviral and/or antiproliferative activities designed for the management of polyomavirus-associated diseases. The severe diseases caused by polyomaviruses, mainly in immunocompromised hosts, and the absence of licensed treatments, make the discovery of new antipolyomavirus drugs urgent. Parallels can be made between the SV40 L-Tag and the human papillomavirus (HPV) oncoproteins (E6 and E7) as they are also able to deregulate the cell cycle in order to promote cell transformation and its maintenance. In this review, a presentation of the SV40 L-Tag characteristics, regarding viral replication and cellular transformation, will show how similar these two processes are between the polyoma- and papillomavirus families. Insights at the molecular level will highlight similarities in the binding of polyoma- and papillomavirus replicative helicases to the viral DNA and in their disruptions of the p53 and pRb tumor suppressor proteins.

  2. Assembly and Purification of Polyomavirus-Like Particles from Plants.

    PubMed

    Catrice, Emeline V B; Sainsbury, Frank

    2015-10-01

    Polyomaviruses are small DNA viruses that have a history of use in biotechnology. The capsids of a number of species have been developed into experimental prophylactic and therapeutic virus-like particle (VLP) vaccines. In order to explore plants as a host for the expression and purification of polyomavirus-like particles, we have transiently expressed the major capsid protein, VP1, in Nicotiana benthamiana leaves. Deletion of a polybasic motif from the N-terminal region of VP1 resulted in increased expression as well as reduced necrosis of leaf tissue, which was associated with differences in subcellular localisation and reduced DNA binding by the deletion variant (ΔVP1). Self-assembled VLPs were recovered from tissue expressing both wild-type VP1 and ΔVP1 by density gradient ultracentrifugation. VLPs composed of ΔVP1 were more homogenous than wtVPLs and, unlike the latter, did not encapsidate nucleic acid. Such homogenous, empty VLPs are of great interest in biotechnology and nanotechnology. In addition, we show that both MPyV VLP variants assembled in plants can be produced with encapsidated foreign protein. Thus, this study demonstrates the utility of plant-based expression of polyomavirus-like particles and the suitability of this host for further developments in polyomavirus-based technologies.

  3. Identification of a novel polyomavirus from vervet monkeys in Zambia.

    PubMed

    Yamaguchi, Hiroki; Kobayashi, Shintaro; Ishii, Akihiro; Ogawa, Hirohito; Nakamura, Ichiro; Moonga, Ladslav; Hang'ombe, Bernard M; Mweene, Aaron S; Thomas, Yuka; Kimura, Takashi; Sawa, Hirofumi; Orba, Yasuko

    2013-06-01

    To examine polyomavirus (PyV) infection in wildlife, we investigated the presence of PyVs in Zambia with permission from the Zambia Wildlife Authority. We analysed 200 DNA samples from the spleens and kidneys (n = 100 each) of yellow baboons and vervet monkeys (VMs) (n = 50 each). We detected seven PyV genome fragments in 200 DNA samples using a nested broad-spectrum PCR method, and identified five full-length viral genomes using an inverse PCR method. Phylogenetic analysis of virally encoded proteins revealed that four PyVs were closely related to either African green monkey PyV or simian agent 12. Only one virus detected from a VM spleen was found to be related, with relatively low nucleotide sequence identity (74 %), to the chimpanzee PyV, which shares 48 % nucleotide sequence identity with the human Merkel cell PyV identified from Merkel cell carcinoma. The obtained entire genome of this virus was 5157 bp and had large T- and small t-antigens, and VP1 and VP2 ORFs. This virus was tentatively named vervet monkey PyV 1 (VmPyV1) as a novel PyV. Comparison with other PyVs revealed that VmPyV1, like chimpanzee PyV, had a longer VP1 ORF. To examine whether the VmPyV1 genome could produce viral proteins in cultured cells, the whole genome was transfected into HEK293T cells. We detected VP1 protein expression in the transfected HEK293T cells by immunocytochemical and immunoblot analyses. Thus, we identified a novel PyV genome from VM spleen.

  4. BK Polyomavirus Genotypes Represent Distinct Serotypes with Distinct Entry Tropism

    PubMed Central

    Pastrana, Diana V.; Ray, Upasana; Magaldi, Thomas G.; Schowalter, Rachel M.; Çuburu, Nicolas

    2013-01-01

    BK polyomavirus (BKV) causes significant urinary tract pathogenesis in immunosuppressed individuals, including kidney and bone marrow transplant recipients. It is currently unclear whether BKV-neutralizing antibodies can moderate or prevent BKV disease. We developed reporter pseudoviruses based on seven divergent BKV isolates and performed neutralization assays on sera from healthy human subjects. The results demonstrate that BKV genotypes I, II, III, and IV are fully distinct serotypes. While nearly all healthy subjects had BKV genotype I-neutralizing antibodies, a majority of subjects did not detectably neutralize genotype III or IV. Surprisingly, BKV subgenotypes Ib1 and Ib2 can behave as fully distinct serotypes. This difference is governed by as few as two residues adjacent to the cellular glycan receptor-binding site on the virion surface. Serological analysis of mice given virus-like particle (VLP)-based BKV vaccines confirmed these findings. Mice administered a multivalent VLP vaccine showed high-titer serum antibody responses that potently cross-neutralized all tested BKV genotypes. Interestingly, each of the neutralization serotypes bound a distinct spectrum of cell surface receptors, suggesting a possible connection between escape from recognition by neutralizing antibodies and cellular attachment mechanisms. The finding implies that different BKV genotypes have different cellular tropisms and pathogenic potentials in vivo. Individuals who are infected with one BKV serotype may remain humorally vulnerable to other BKV serotypes after implementation of T cell immunosuppression. Thus, prevaccinating organ transplant recipients with a multivalent BKV VLP vaccine might reduce the risk of developing posttransplant BKV disease. PMID:23843634

  5. Neutralization Serotyping of BK Polyomavirus Infection in Kidney Transplant Recipients

    PubMed Central

    Pastrana, Diana V.; Brennan, Daniel C.; Çuburu, Nicolas; Storch, Gregory A.; Viscidi, Raphael P.; Randhawa, Parmjeet S.; Buck, Christopher B.

    2012-01-01

    BK polyomavirus (BKV or BKPyV) associated nephropathy affects up to 10% of kidney transplant recipients (KTRs). BKV isolates are categorized into four genotypes. It is currently unclear whether the four genotypes are also serotypes. To address this issue, we developed high-throughput serological assays based on antibody-mediated neutralization of BKV genotype I and IV reporter vectors (pseudoviruses). Neutralization-based testing of sera from mice immunized with BKV-I or BKV-IV virus-like particles (VLPs) or sera from naturally infected human subjects revealed that BKV-I specific serum antibodies are poorly neutralizing against BKV-IV and vice versa. The fact that BKV-I and BKV-IV are distinct serotypes was less evident in traditional VLP-based ELISAs. BKV-I and BKV-IV neutralization assays were used to examine BKV type-specific neutralizing antibody responses in KTRs at various time points after transplantation. At study entry, sera from 5% and 49% of KTRs showed no detectable neutralizing activity for BKV-I or BKV-IV neutralization, respectively. By one year after transplantation, all KTRs were neutralization seropositive for BKV-I, and 43% of the initially BKV-IV seronegative subjects showed evidence of acute seroconversion for BKV-IV neutralization. The results suggest a model in which BKV-IV-specific seroconversion reflects a de novo BKV-IV infection in KTRs who initially lack protective antibody responses capable of neutralizing genotype IV BKVs. If this model is correct, it suggests that pre-vaccinating prospective KTRs with a multivalent VLP-based vaccine against all BKV serotypes, or administration of BKV-neutralizing antibodies, might offer protection against graft loss or dysfunction due to BKV associated nephropathy. PMID:22511874

  6. 5-HT2 receptors facilitate JC polyomavirus entry.

    PubMed

    Assetta, Benedetta; Maginnis, Melissa S; Gracia Ahufinger, Irene; Haley, Sheila A; Gee, Gretchen V; Nelson, Christian D S; O'Hara, Bethany A; Allen Ramdial, Stacy-ann A; Atwood, Walter J

    2013-12-01

    The human JC polyomavirus (JCPyV) causes the rapidly progressing demyelinating disease progressive multifocal leukoencephalopathy (PML). The disease occurs most often in individuals with AIDS but also occurs in individuals receiving immunomodulatory therapies for immune-related diseases such as multiple sclerosis. JCPyV infection of host cells requires the pentasaccharide lactoseries tetrasaccharide c (LSTc) and the serotonin receptor 5-hydroxytryptamine (5-HT) receptor 5-HT2AR. While LSTc is involved in the initial attachment of virus to cells via interactions with VP1, the mechanism by which 5-HT2AR contributes to infection is not clear. To further define the roles of serotonin receptors in infection, HEK293A cells, which are poorly permissive to JCPyV, were transfected with 14 different isoforms of serotonin receptor. Only 5-HT2 receptors were found to support infection by JCPyV. None of the other 11 isoforms of serotonin receptor supported JCPyV infection. Expression of 5-HT2 receptors did not increase binding of JCPyV to cells, but this was not unexpected, given that the cells uniformly expressed the major attachment receptor, LSTc. Infection of these cells remained sensitive to inhibition with soluble LSTc, confirming that LSTc recognition is required for JCPyV infection. Virus internalization into HEK293A cells was significantly and specifically enhanced when 5HT2 receptors were expressed. Taken together, these data confirm that the carbohydrate LSTc is the attachment receptor for JCPyV and that the type 2 serotonin receptors contribute to JCPyV infection by facilitating entry.

  7. IFPA Meeting 2013 Workshop Report III: maternal placental immunological interactions, novel determinants of trophoblast cell fate, dual ex vivo perfusion of the human placenta.

    PubMed

    Abumaree, M H; Brownbill, P; Burton, G; Castillo, C; Chamley, L; Croy, B A; Drewlo, S; Dunk, C; Girard, S; Hansson, S; Jones, S; Jurisicova, A; Lewis, R; Letarte, M; Parast, M; Pehrson, C; Rappolee, D; Schneider, H; Tannetta, D; Varmuza, S; Wadsack, C; Wallace, A E; Zenerino, C; Lash, G E

    2014-02-01

    Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialised topics. At IFPA meeting 2013 there were twelve themed workshops, three of which are summarized in this report. These workshops related to various aspects of placental biology but collectively covered areas of placental function, cell turnover and immunology: 1) immunology; 2) novel determinants of placental cell fate; 3) dual perfusion of human placental tissue.

  8. IFPA Meeting 2012 Workshop Report I: comparative placentation and animal models, advanced techniques in placental histopathology, human pluripotent stem cells as a model for trophoblast differentiation.

    PubMed

    Ackerman, W E; Carter, A M; De Mestre, A M; Golos, T G; Jeschke, U; Kusakabe, K; Laurent, L C; Parast, M M; Roberts, R M; Robinson, J M; Rutherford, J; Soma, H; Takizawa, T; Ui-Tei, K; Lash, G E

    2013-03-01

    Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialized topics. At IFPA meeting 2012 there were twelve themed workshops, three of which are summarized in this report. These workshops related to various aspects of placental biology but collectively covered areas of models and technical issues involved in placenta research: 1) comparative placentation and animal models; 2) advanced techniques in placental histopathology; 3) human pluripotent stem cells as a model for trophoblast differentiation.

  9. Summary Report: State-of-the-Science Workshop on Chemically-Induced Mouse Lung Tumors: Applications to Human Health Assessments

    EPA Science Inventory

    The EPA hosted a two-day, state-of-the-science workshop which covered a broad range of evidence from human, animal, and in vitro studies with a focus on specific chemicals (ethylbenzene, naphthalene, and styrene) that cause lung tumors in mice and are implicated in a proposed spe...

  10. Human health risk assessment of lead, manganese and copper from scrapped car paint dust from automobile workshops in Nigeria.

    PubMed

    Nduka, John Kanayochukwu; Onyenezi Amuka, John Paul; Onwuka, Jude Chinedu; Udowelle, Nnaemeka Arinze; Orisakwe, Orish Ebere

    2016-10-01

    The economic downturn in Nigeria and Structural Adjustment Programme led to the flooding of Nigerian market with imported used automobiles. Most of these vehicles needed refurbishing and reworking. The present study is a human health risk assessment of metal exposure resulting from reworking of imported used vehicles in Nigeria. Scrap paint dusts from 56 Japanese made cars were collected from 8 different mechanic villages (workshops A-H] in Southeastern Nigeria. Scrap paints were homogenized, mixed, divided into fine particles and digested by standard method. The filtrates were assayed of lead, manganese and copper with atomic absorption spectrophotometry (AAS). Workshop B has the highest concentration of Pb (4.26 ± 0.93). Manganese in workshops A and F were (3.31 ± 0.85) and (3.04 ± 0.47) respectively and were higher than the levels from workshops C, B, D, G and H. Copper in workshop D (7.11 ± 0.21) was significantly greater than the other workshops. The highest hazard quotient (HQ) through ingestion, inhalation and dermal exposures in adults were 9.44E-05 (workshop B), 4.20E-01 (workshop B) and 1.08E-05 (workshop D) respectively. The highest values for HQ through ingestion, inhalation and dermal in children were 8.82E-04, 7.61E-01 and 2.86E-05 all in workshop B respectively. For children, the highest carcinogenic risk levels were 7.05E-08, 6.09E-05 and 2.29E-10 for ingestion, inhalation and dermal exposures respectively. In adults, the carcinogenic risk levels were 7.55E-09, 3.39E-05 and 8.67E-10 for ingestion, inhalation and dermal exposures respectively. Chronic exposure to scrap car paint dusts may be of significant public health importance in Nigeria as this may add to the body burden of some heavy metals.

  11. Bromodomain Protein Brd4 Plays a Key Role in Merkel Cell Polyomavirus DNA Replication

    PubMed Central

    Wang, Xin; Li, Jing; Schowalter, Rachel M.; Jiao, Jing; Buck, Christopher B.; You, Jianxin

    2012-01-01

    Merkel cell polyomavirus (MCV or MCPyV) is the first human polyomavirus to be definitively linked to cancer. The mechanisms of MCV-induced oncogenesis and much of MCV biology are largely unexplored. In this study, we demonstrate that bromodomain protein 4 (Brd4) interacts with MCV large T antigen (LT) and plays a critical role in viral DNA replication. Brd4 knockdown inhibits MCV replication, which can be rescued by recombinant Brd4. Brd4 colocalizes with the MCV LT/replication origin complex in the nucleus and recruits replication factor C (RFC) to the viral replication sites. A dominant negative inhibitor of the Brd4-MCV LT interaction can dissociate Brd4 and RFC from the viral replication complex and abrogate MCV replication. Furthermore, obstructing the physiologic interaction between Brd4 and host chromatin with the chemical compound JQ1(+) leads to enhanced MCV DNA replication, demonstrating that the role of Brd4 in MCV replication is distinct from its role in chromatin-associated transcriptional regulation. Our findings demonstrate mechanistic details of the MCV replication machinery; providing novel insight to elucidate the life cycle of this newly discovered oncogenic DNA virus. PMID:23144621

  12. Reducing persistent polyomavirus infection increases functionality of virus-specific memory CD8 T cells.

    PubMed

    Qin, Qingsong; Lauver, Matthew; Maru, Saumya; Lin, Eugene; Lukacher, Aron E

    2017-02-01

    Mouse polyomavirus (MuPyV) causes a smoldering persistent infection in immunocompetent mice. To lower MuPyV infection in acutely and persistently infected mice, and study the impact of a temporal reduction in viral loads on the memory CD8 T cell response, we created a recombinant MuPyV in which a loxP sequence was inserted into the A2 strain genome upstream of the early promoter and another loxP sequence was inserted in cis into the intron shared by all three T antigens. Using mice transgenic for tamoxifen-inducible Cre recombinase, we demonstrated that reduction in MuPyV load during persistent infection was associated with differentiation of virus-specific CD8 T cells having a superior recall response. Evidence presented here supports the concept that reduction in viral load during persistent infection can promote differentiation of protective virus-specific memory CD8 T cells in patients at risk for diseases caused by human polyomaviruses.

  13. Summary Report: Workshop on the Potential Risks of Antibody-Dependent Enhancement in Human HIV Vaccine Trials

    DTIC Science & Technology

    1993-01-01

    Gidlund M , Chiodi F, et al.: Enhancement of human of gag genes from 70 international HIV- I isolates provides evi- immunodeficiency virus (HIV...Workshop on the Potential Risks of Antibody-Dependent Enhancement in Human HIV Vaccine Trials JOHN R. MASCOLA,’ BONNIE J. MATHIESON, 2 PHILIP M . ZACK...monocyte/macrophage ( M / M ) cells is the pathophysiological Army Institute of Research and was hosted by the Health Sciences mechanism for enhanced disease

  14. Human-based approaches to pharmacology and cardiology: an interdisciplinary and intersectorial workshop

    PubMed Central

    Rodriguez, Blanca; Carusi, Annamaria; Abi-Gerges, Najah; Ariga, Rina; Britton, Oliver; Bub, Gil; Bueno-Orovio, Alfonso; Burton, Rebecca A.B.; Carapella, Valentina; Cardone-Noott, Louie; Daniels, Matthew J.; Davies, Mark R.; Dutta, Sara; Ghetti, Andre; Grau, Vicente; Harmer, Stephen; Kopljar, Ivan; Lambiase, Pier; Lu, Hua Rong; Lyon, Aurore; Minchole, Ana; Muszkiewicz, Anna; Oster, Julien; Paci, Michelangelo; Passini, Elisa; Severi, Stefano; Taggart, Peter; Tinker, Andy; Valentin, Jean-Pierre; Varro, Andras; Wallman, Mikael; Zhou, Xin

    2016-01-01

    Both biomedical research and clinical practice rely on complex datasets for the physiological and genetic characterization of human hearts in health and disease. Given the complexity and variety of approaches and recordings, there is now growing recognition of the need to embed computational methods in cardiovascular medicine and science for analysis, integration and prediction. This paper describes a Workshop on Computational Cardiovascular Science that created an international, interdisciplinary and inter-sectorial forum to define the next steps for a human-based approach to disease supported by computational methodologies. The main ideas highlighted were (i) a shift towards human-based methodologies, spurred by advances in new in silico, in vivo, in vitro, and ex vivo techniques and the increasing acknowledgement of the limitations of animal models. (ii) Computational approaches complement, expand, bridge, and integrate in vitro, in vivo, and ex vivo experimental and clinical data and methods, and as such they are an integral part of human-based methodologies in pharmacology and medicine. (iii) The effective implementation of multi- and interdisciplinary approaches, teams, and training combining and integrating computational methods with experimental and clinical approaches across academia, industry, and healthcare settings is a priority. (iv) The human-based cross-disciplinary approach requires experts in specific methodologies and domains, who also have the capacity to communicate and collaborate across disciplines and cross-sector environments. (v) This new translational domain for human-based cardiology and pharmacology requires new partnerships supported financially and institutionally across sectors. Institutional, organizational, and social barriers must be identified, understood and overcome in each specific setting. PMID:26622055

  15. Human-based approaches to pharmacology and cardiology: an interdisciplinary and intersectorial workshop.

    PubMed

    Rodriguez, Blanca; Carusi, Annamaria; Abi-Gerges, Najah; Ariga, Rina; Britton, Oliver; Bub, Gil; Bueno-Orovio, Alfonso; Burton, Rebecca A B; Carapella, Valentina; Cardone-Noott, Louie; Daniels, Matthew J; Davies, Mark R; Dutta, Sara; Ghetti, Andre; Grau, Vicente; Harmer, Stephen; Kopljar, Ivan; Lambiase, Pier; Lu, Hua Rong; Lyon, Aurore; Minchole, Ana; Muszkiewicz, Anna; Oster, Julien; Paci, Michelangelo; Passini, Elisa; Severi, Stefano; Taggart, Peter; Tinker, Andy; Valentin, Jean-Pierre; Varro, Andras; Wallman, Mikael; Zhou, Xin

    2016-09-01

    Both biomedical research and clinical practice rely on complex datasets for the physiological and genetic characterization of human hearts in health and disease. Given the complexity and variety of approaches and recordings, there is now growing recognition of the need to embed computational methods in cardiovascular medicine and science for analysis, integration and prediction. This paper describes a Workshop on Computational Cardiovascular Science that created an international, interdisciplinary and inter-sectorial forum to define the next steps for a human-based approach to disease supported by computational methodologies. The main ideas highlighted were (i) a shift towards human-based methodologies, spurred by advances in new in silico, in vivo, in vitro, and ex vivo techniques and the increasing acknowledgement of the limitations of animal models. (ii) Computational approaches complement, expand, bridge, and integrate in vitro, in vivo, and ex vivo experimental and clinical data and methods, and as such they are an integral part of human-based methodologies in pharmacology and medicine. (iii) The effective implementation of multi- and interdisciplinary approaches, teams, and training combining and integrating computational methods with experimental and clinical approaches across academia, industry, and healthcare settings is a priority. (iv) The human-based cross-disciplinary approach requires experts in specific methodologies and domains, who also have the capacity to communicate and collaborate across disciplines and cross-sector environments. (v) This new translational domain for human-based cardiology and pharmacology requires new partnerships supported financially and institutionally across sectors. Institutional, organizational, and social barriers must be identified, understood and overcome in each specific setting.

  16. Reporting of the third international workshop on human chromosome 22 mapping

    SciTech Connect

    Emanuel, B.S.; Buetow, K.; Nussbaum, R.; Scambler, P; Lipinski, M.; Overton, C.

    1992-12-31

    The third international workshop on the mapping of human chromosome 22 was held at the Sugarloaf Conference Center in Philadelphia Pennsylvania USA from September 17--20, 1992. It was organized by Beverly S. Emanuel of Children`s Hospital of Philadelphia and the Human Genome Center for Chromosome 22. The highlights of the conference included the discussion of the chromosome 22 gene at the Ewings Sarcoma breakpoint, the identification of a polymorphic TG/CA repeat containing locus tightly linked to the NF2 gene, the isolation of a candidate tumor suppressor locus for meningioma, the isolation of numerous as yet uncharacterized new cDNAs for chromosome 22 and the progress which has been made on generating physical and genetic maps of the chromosome. There is a new genetic map comprised of 16 short tandem repeat polymorphism (STRP) markers of which 12 have greater than 70% heterozygosity (Fig. 1). It was decided that the next meeting should be held in 18 months and it will be organized by Peter Scambler in the United Kingdom.

  17. BK Polyomavirus and the Transplanted Kidney: Immunopathology and Therapeutic Approaches

    PubMed Central

    Lamarche, Caroline; Orio, Julie; Collette, Suzon; Senécal, Lynne; Hébert, Marie-Josée; Renoult, Édith; Tibbles, Lee Anne; Delisle, Jean-Sébastien

    2016-01-01

    Abstract BK polyomavirus is ubiquitous, with a seropositivity rate of over 75% in the adult population. Primary infection is thought to occur in the respiratory tract, but asymptomatic BK virus latency is established in the urothelium. In immunocompromised host, the virus can reactivate but rarely compromises kidney function except in renal grafts, where it causes a tubulointerstitial inflammatory response similar to acute rejection. Restoring host immunity against the virus is the cornerstone of treatment. This review covers the virus-intrinsic features, the posttransplant microenvironment as well as the host immune factors that underlie the pathophysiology of polyomavirus-associated nephropathy. Current and promising therapeutic approaches to treat or prevent this complication are discussed in relation to the complex immunopathology of this condition. PMID:27391196

  18. Complete Genome Sequence of a Polyomavirus Recovered from a Pomona Leaf-Nosed Bat (Hipposideros pomona) Metagenome Data Set

    PubMed Central

    Cantalupo, Paul G.; Buck, Christopher B.

    2017-01-01

    ABSTRACT We report here the complete genome sequence of a polyomavirus found in a nasal/rectal metagenome of Hipposideros pomona (Pomona leaf-nosed bat). Interestingly, the genetic organization and phylogenetic relationships of the new virus suggest greater similarity to recently discovered fish-associated polyomaviruses rather than to polyomavirus species previously observed in bats. PMID:28104645

  19. Genomic characterization of two novel polyomaviruses in Brazilian insectivorous bats.

    PubMed

    de Sales Lima, Francisco Esmaile; Cibulski, Samuel Paulo; Witt, André Alberto; Franco, Ana Cláudia; Roehe, Paulo Michel

    2015-07-01

    Two novel genomes comprising ≈4.9 kb were identified by next-generation sequencing from pooled organs of Tadarida brasiliensis bats. The overall nucleotide sequence identities between the viral genomes characterized here were less than 80% in comparison to other polyomaviruses (PyVs), members of the family Polyomaviridae. The new genomes display the archetypal organization of PyVs, which includes open reading frames for the regulatory proteins small T antigen (STAg) and large T antigen (LTAg), as well as capsid proteins VP1, VP2 and VP3. In addition, an alternate ORF was identified in the early genome region that is conserved in a large monophyletic group of polyomaviruses. Phylogenetic analysis showed similar clustering with group of PyVs detected in Otomops and Chaerephon bats and some species of monkeys. In this study, the genomes of two novel PyVs were detected in bats of a single species, demonstrating that these mammals can harbor genetically diverse polyomaviruses.

  20. Specific IgG Antibodies React to Mimotopes of BK Polyomavirus, a Small DNA Tumor Virus, in Healthy Adult Sera

    PubMed Central

    Pietrobon, Silvia; Bononi, Ilaria; Mazzoni, Elisa; Lotito, Francesca; Manfrini, Marco; Puozzo, Andrea; Destro, Federica; Guerra, Giovanni; Nocini, Pier Francesco; Martini, Fernanda; Tognon, Mauro G.

    2017-01-01

    BK polyomavirus (BKPyV) was isolated in 1971 from the urine of a kidney transplant patient. Soon after its identification, BKPyV was characterized as a kidney-tropic virus, which is responsible of a significant fraction of the rejection of transplant kidney in the host. Moreover, in experimental conditions, BKPyV is able to transform different types of animal and human cells and to induce tumors of different histotypes in experimental animals. BKPyV DNA sequences have been detected in healthy individuals and cancer patients using polymerase chain reaction/Shouthern blot hybridization methods. Serum antibodies against this polyomavirus were revealed using immunological techniques, which, however, cross-react with other polyomaviruses such as JC (JCPyV) and Simian Virus 40. These non-specific data indicate the need of novel immunological methods and new investigations to check in a specific manner, BKPyV spread in humans. To this aim, mimotopes from BKPyV structural capsid protein 1 (VP1) were employed for specific immunological reactions to IgG antibodies of human serum samples. An indirect enzyme-linked immunosorbent assay with synthetic peptides mimicking immunogenic epitopes of BKPyV VP1 was set up and employed to test sera of healthy adult subjects. Data from this innovative immunological assay indicate that serum antibodies against BKPyV VP1 mimotopes are detectable in healthy subjects ranging from 18 to 90 years old. The overall prevalence of serum samples that reacted to BKPyV VP1 mimotopes was 72%. The strong points from this investigation are the novelty of the immunological method, its simplicity of the approach, and the specificity of BKPyV antibody reaction to VP1 mimotopes. PMID:28321224

  1. Trichodysplasia spinulosa: a polyomavirus infection specifically targeting follicular keratinocytes in immunocompromised patients.

    PubMed

    Rouanet, J; Aubin, F; Gaboriaud, P; Berthon, P; Feltkamp, M C; Bessenay, L; Touzé, A; Nicol, J T J; Franck, F; D'Incan, M

    2016-03-01

    Trichodysplasia spinulosa (TS) is a rare skin disease, caused by a specific polyomavirus, occurring in immunocompromised patients. The pathophysiological mechanisms of TS are not yet fully understood. By using polymerase chain reaction and skin biopsy immunostaining we report evidence, in a paediatric case, of follicular keratinocytes being the primary target of trichodysplasia spinulosa-associated polyomavirus.

  2. Diagnostic methods for and clinical pictures of polyomavirus primary infections in children, Finland.

    PubMed

    Chen, Tingting; Tanner, Laura; Simell, Ville; Hedman, Lea; Mäkinen, Marjaana; Sadeghi, Mohammadreza; Veijola, Riitta; Hyöty, Heikki; Ilonen, Jorma; Knip, Mikael; Toppari, Jorma; Simell, Olli; Söderlund-Venermo, Maria; Hedman, Klaus

    2014-04-01

    We used comprehensive serodiagnostic methods (IgM, IgG, and IgG avidity) and PCR to study Merkel cell polyomavirus and trichodysplasia spinulosa-associated polyomavirus infections in children observed from infancy to adolescence. Comparing seroconversion intervals with previous and subsequent intervals, we found that primary infections with these 2 viruses were asymptomatic in childhood.

  3. Characterization of novel polyomaviruses from Bornean and Sumatran orang-utans.

    PubMed

    Groenewoud, Marlous J; Fagrouch, Zahra; van Gessel, Sabine; Niphuis, Henk; Bulavaite, Aiste; Warren, Kristin S; Heeney, Jonathan L; Verschoor, Ernst J

    2010-03-01

    Serological screening of sera from orang-utans demonstrated a high percentage of sera that cross-reacted with antigens of the polyomavirus (PyV) simian virus 40. Analysis of archival DNA samples from 71 Bornean and eight Sumatran orang-utans with a broad-spectrum PCR assay resulted in the detection of PyV infections in 11 animals from both species. Sequence analysis of the amplicons revealed considerable differences between the PyVs from Bornean and Sumatran orang-utans. The genome from two PyVs, one from each species, was therefore amplified and sequenced. Both viral genomes revealed a characteristic PyV architecture, but lacked an obvious agnogene. Neighbour-joining analysis positioned the viruses in a large cluster together with viruses from bats, bovines, rodents and several primate PyVs from chimpanzees, African green monkeys, squirrel monkeys and the human Merkel cell PyV.

  4. Characterization of Monoclonal Antibodies Specific for the Merkel Cell Polyomavirus Capsid

    PubMed Central

    Pastrana, Diana V.; Pumphrey, Katherine A.; Çuburu, Nicolas; Schowalter, Rachel M.; Buck, Christopher B.

    2010-01-01

    Merkel cell polyomavirus (MCV) has been implicated as a causative agent in Merkel cell carcinoma. Robust polyclonal antibody responses against MCV have been documented in human subjects, but monoclonal antibodies (mAbs) specific for the VP1 capsid protein have not yet been characterized. We generated 12 mAbs capable of binding recombinant MCV virus-like particles. The use of a short immunogenic priming schedule was important for production of the mAbs. Ten of the 12 mAbs were highly effective for immunofluorescent staining of cells expressing capsid proteins. An overlapping set of 10 mAbs were able to neutralize the infectivity of MCV-based reporter vectors, with 50% effective doses in the low picomolar range. Three mAbs interfered with the binding of MCV virus-like particles to cells. This panel of anti-capsid antibodies should provide a useful set of tools for the study of MCV. PMID:20598728

  5. In Vitro Replication Assay for Merkel Cell Polyomavirus (MCPyV).

    PubMed

    Neumann, Friederike; Czech-Sioli, Manja; Grundhoff, Adam; Fischer, Nicole

    2015-08-03

    Merkel cell polyomavirus (MCPyV) genomes are clonally integrated in tumor cells of ∼95% of all Merkel cell carcinoma (MCC) cases. The virus is highly prevalent; however, where the virus persists and which cell types are permissive for MCPyV replication is still unknown. As a consequence, very little information is available about the life cycle and no fully permissive in vitro replication system has been established. Recently, semi-permissive replication systems based on wild-type MCPyV genomes recovered from the skin of healthy donors or synthetic MCPyV genomes constructed from consensus sequences have been established. The transfection of this intramolecular re-circularized MCPyV DNA into some human cell lines recapitulates efficient DNA replication of the viral genome, viral gene expression as well as moderate levels of virus particle formation. However, serial transmission of infectious virus is still restricted in these cells.

  6. Butcherbird polyomavirus isolated from a grey butcherbird (Cracticus torquatus) in Queensland, Australia.

    PubMed

    Bennett, Mark D; Gillett, Amber

    2014-01-31

    A novel avian polyomavirus was detected in peri-ocular skin lesions collected from a grey butcherbird (Cracticus torquatus), using a combination of multiply primed rolling circle amplification, nested PCR and long range PCR. The sequence of Butcherbird polyomavirus was determined by combining next generation sequencing and primer walking techniques. The circular double-stranded DNA genome of Butcherbird polyomavirus consisted of 5084 bp, and encoded six open reading frames (ORF-X, VP2, VP3, VP1, small T-antigen and large T-antigen). Phylogenetic analysis placed it amongst other members of the genus Avipolyomavirus, most closely related to Crow polyomavirus. Next generation sequencing enabled the detection of DNA fragments similar to, but distinct from, Canarypox virus within the same lesion from which Butcherbird polyomavirus was amplified, thus confirming an avipolyomavirus-avipoxvirus co-infection in the peri-ocular skin lesions of this grey butcherbird.

  7. Report of the first international workshop on human chromosome 8 mapping. Final report

    SciTech Connect

    Wood, S.; Ben Othmane, K.; Bergerheim, U.S.R.

    1993-12-31

    The first international chromosome 8 workshop was held in Vancouver, Canada May 2--4, 1993. The conference was attended by 23 participants from Australia, Canada, Germany, the Netherlands, Sweden, the United Kingdom and the US. Twenty three abstracts are included from this workshop. The workshop was supported by CGAT/CTAG (Canadian Genome Analysis & Technology Program/Programme Canadien de Technologie & D`Analyse du Genome) as well as by travel funds allocated by the National Institutes of Health and the Department of Energy of the United States and by agencies within the countries of overseas participants. The goals of the workshop were to evaluate new locus assignments, review new data obtained for previously assigned loci, develop a consensus marker order for chromosome 8, assess and integrate physical mapping information, identify resources and foster collaboration.

  8. Discovery of a polyomavirus in European badgers (Meles meles) and the evolution of host range in the family Polyomaviridae.

    PubMed

    Hill, Sarah C; Murphy, Aisling A; Cotten, Matthew; Palser, Anne L; Benson, Phillip; Lesellier, Sandrine; Gormley, Eamonn; Richomme, Céline; Grierson, Sylvia; Bhuachalla, Deirdre Ni; Chambers, Mark; Kellam, Paul; Boschiroli, María-Laura; Ehlers, Bernhard; Jarvis, Michael A; Pybus, Oliver G

    2015-06-01

    Polyomaviruses infect a diverse range of mammalian and avian hosts, and are associated with a variety of symptoms. However, it is unknown whether the viruses are found in all mammalian families and the evolutionary history of the polyomaviruses is still unclear. Here, we report the discovery of a novel polyomavirus in the European badger (Meles meles), which to our knowledge represents the first polyomavirus to be characterized in the family Mustelidae, and within a European carnivoran. Although the virus was discovered serendipitously in the supernatant of a cell culture inoculated with badger material, we subsequently confirmed its presence in wild badgers. The European badger polyomavirus was tentatively named Meles meles polyomavirus 1 (MmelPyV1). The genome is 5187 bp long and encodes proteins typical of polyomaviruses. Phylogenetic analyses including all known polyomavirus genomes consistently group MmelPyV1 with California sea lion polyomavirus 1 across all regions of the genome. Further evolutionary analyses revealed phylogenetic discordance amongst polyomavirus genome regions, possibly arising from evolutionary rate heterogeneity, and a complex association between polyomavirus phylogeny and host taxonomic groups.

  9. 77 FR 12313 - Food Labeling Workshop; Public Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-29

    ... HUMAN SERVICES Food and Drug Administration Food Labeling Workshop; Public Workshop AGENCY: Food and Drug Administration, HHS. ACTION: Notice of public workshop. The Food and Drug Administration (FDA... University (OSU), Robert M. Kerr Food & Agricultural Products Center (FAPC), is announcing a public...

  10. Workshop 7 (synthesis): towards a recycling society: systems approach to small-scale reuse of human waste.

    PubMed

    Rahman, A; Drangert, J O

    2001-01-01

    Population increase and growing quantities of human excreta create serious problems and high risks for people's health. Alternative solutions are becoming crucial to improve urban living conditions, and to shorten water and nutrient flows into circulation of used water and nutrient in human excreta. The speakers presented a wide range of experiences of "closing the loops" and thereby turning potential waste into productive use. The focus of the workshop deliberations was on simplifying the hygienisation of urine and faeces and the reuse in food production by using urine-diverting toilets, as well as innovative ways to recycle sewage after various stages of treatment.

  11. CRISPR/Cas9 System as an Agent for Eliminating Polyomavirus JC Infection

    PubMed Central

    Wollebo, Hassen S.; Bellizzi, Anna; Kaminski, Rafal; Hu, Wenhui; White, Martyn K.; Khalili, Kamel

    2015-01-01

    Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease of the central nervous system (CNS) caused by reactivation of the human polyomavirus JCV gene expression and its replication in oligodendrocytes, the myelin producing cells in the brain. Once a rare disease seen in patients with lymphotproliferative and myeloproliferative disorders, PML has been seen more frequently in HIV-1 positive/AIDS patients as well as patients undergoing immunomodulatory therapy due for autoimmune disorders including multiple sclerosis, rheumatoid arthritis, and others. As of now there is no cure for PML and in most cases disease progression leads to death within two years. Similar to other polyomaviruses, the JCV genome is small circular double stranded DNA that includes coding sequences for the viral early protein, T-antigen, which is critical for directing viral reactivation and lytic infection. Here, we employ a newly developed gene editing strategy, CRISPR/Cas9, to introduce mutations in the viral genome and, by inactivating the gene encoding T-antigen, inhibit viral replication. We first used bioinformatics screening and identified several potential targets within the JCV T-antigen gene that can serve as sites for the creation of guide RNAs (gRNAs) for positioning the Cas9 nuclease on the designated area of the viral genome for editing. Results from a series of integrated genetic and functional studies showed that transient or conditional expression of Cas9 and gRNAs specifically targets the DNA sequences corresponding to the N-terminal region of T-antigen, and by introducing mutation, interferes with expression and function of of the viral protein, hence suppressing viral replication in permissive cells. Results from SURVEYOR assay revealed no off-target effects of the JCV-specific CRISPR/Cas9 editing apparatus. These observations provide the first evidence for the employment of a gene editing strategy as a promising tool for the elimination of the JCV

  12. CRISPR/Cas9 System as an Agent for Eliminating Polyomavirus JC Infection.

    PubMed

    Wollebo, Hassen S; Bellizzi, Anna; Kaminski, Rafal; Hu, Wenhui; White, Martyn K; Khalili, Kamel

    2015-01-01

    Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease of the central nervous system (CNS) caused by reactivation of the human polyomavirus JCV gene expression and its replication in oligodendrocytes, the myelin producing cells in the brain. Once a rare disease seen in patients with lymphotproliferative and myeloproliferative disorders, PML has been seen more frequently in HIV-1 positive/AIDS patients as well as patients undergoing immunomodulatory therapy due for autoimmune disorders including multiple sclerosis, rheumatoid arthritis, and others. As of now there is no cure for PML and in most cases disease progression leads to death within two years. Similar to other polyomaviruses, the JCV genome is small circular double stranded DNA that includes coding sequences for the viral early protein, T-antigen, which is critical for directing viral reactivation and lytic infection. Here, we employ a newly developed gene editing strategy, CRISPR/Cas9, to introduce mutations in the viral genome and, by inactivating the gene encoding T-antigen, inhibit viral replication. We first used bioinformatics screening and identified several potential targets within the JCV T-antigen gene that can serve as sites for the creation of guide RNAs (gRNAs) for positioning the Cas9 nuclease on the designated area of the viral genome for editing. Results from a series of integrated genetic and functional studies showed that transient or conditional expression of Cas9 and gRNAs specifically targets the DNA sequences corresponding to the N-terminal region of T-antigen, and by introducing mutation, interferes with expression and function of of the viral protein, hence suppressing viral replication in permissive cells. Results from SURVEYOR assay revealed no off-target effects of the JCV-specific CRISPR/Cas9 editing apparatus. These observations provide the first evidence for the employment of a gene editing strategy as a promising tool for the elimination of the JCV

  13. Early identification of renal transplant recipients with high risk of polyomavirus-associated nephropathy.

    PubMed

    Teutsch, K; Schweitzer, F; Knops, E; Kaiser, R; Pfister, H; Verheyen, J; Göbel, H; Cingöz, T; Di Cristanziano, V

    2015-12-01

    Polyomavirus BK (BKPyV) is ubiquitous among humans. Following primary infection, the virus remains latent predominantly in the hosts' uroepithelial cells. Up to 10 % of renal transplant recipients show a viral reactivation that can lead to polyomavirus-associated nephropathy (PyVAN). In the absence of early treatments, the risk of graft loss is up to 80 %. Monitoring viral load in urine and plasma by real-time PCR after transplantation is the most common diagnostic tool to detect viral reactivation. In the present retrospective study, BKPyV-DNA loads in urine and plasma by quantitative real-time PCR were associated with clinical data, including HLA haplotype, blood parameters and viral genotype, of 40 renal transplant recipients at the University Clinics of Cologne. Seventeen out of 329 patients screened for BKPyV from January 2009 to October 2013 were detected BKPyV positive in urine only, whereas in 23 patients the virus became additionally detectable in plasma. Among these, ten patients progressed to PyVAN. Overall, the present study showed that the detection from the third month onwards after transplantation of a first viruric episode with a median viral load of 1 × 10(8) copies/mL, followed after few days by a first viremic episode with a median viral load of >1 × 10(4) copies/mL, was strongly associated with the development of PyVAN. In conclusion, the viral load and the temporal profile of the first viruric and viremic episode post-transplantation, in combination with specific features of the host immune response, should be considered as relevant clinical determinants of the risk of renal transplant recipients to progress to PyVAN.

  14. Advanced Technologies for Robotic Exploration Leading to Human Exploration: Results from the SpaceOps 2015 Workshop

    NASA Technical Reports Server (NTRS)

    Lupisella, Mark L.; Mueller, Thomas

    2016-01-01

    This paper will provide a summary and analysis of the SpaceOps 2015 Workshop all-day session on "Advanced Technologies for Robotic Exploration, Leading to Human Exploration", held at Fucino Space Center, Italy on June 12th, 2015. The session was primarily intended to explore how robotic missions and robotics technologies more generally can help lead to human exploration missions. The session included a wide range of presentations that were roughly grouped into (1) broader background, conceptual, and high-level operations concepts presentations such as the International Space Exploration Coordination Group Roadmap, followed by (2) more detailed narrower presentations such as rover autonomy and communications. The broader presentations helped to provide context and specific technical hooks, and helped lay a foundation for the narrower presentations on more specific challenges and technologies, as well as for the discussion that followed. The discussion that followed the presentations touched on key questions, themes, actions and potential international collaboration opportunities. Some of the themes that were touched on were (1) multi-agent systems, (2) decentralized command and control, (3) autonomy, (4) low-latency teleoperations, (5) science operations, (6) communications, (7) technology pull vs. technology push, and (8) the roles and challenges of operations in early human architecture and mission concept formulation. A number of potential action items resulted from the workshop session, including: (1) using CCSDS as a further collaboration mechanism for human mission operations, (2) making further contact with subject matter experts, (3) initiating informal collaborative efforts to allow for rapid and efficient implementation, and (4) exploring how SpaceOps can support collaboration and information exchange with human exploration efforts. This paper will summarize the session and provide an overview of the above subjects as they emerged from the SpaceOps 2015

  15. Role of the C-Terminal Region of Vervet Monkey Polyomavirus 1 VP1 in Virion Formation

    PubMed Central

    YAMAGUCHI, Hiroki; KOBAYASHI, Shintaro; MARUYAMA, Junki; SASAKI, Michihito; TAKADA, Ayato; KIMURA, Takashi; SAWA, Hirofumi; ORBA, Yasuko

    2014-01-01

    ABSTRACT Recently, we detected novel vervet monkey polyomavirus 1 (VmPyV) in a vervet monkey. Among amino acid sequences of major capsid protein VP1s of other polyomaviruses, VmPyV VP1 is the longest with additional amino acid residues in the C-terminal region. To examine the role of VmPyV VP1 in virion formation, we generated virus-like particles (VLPs) of VmPyV VP1, because VLP is a useful tool for the investigation of the morphological characters of polyomavirus virions. After the full-length VmPyV VP1 was subcloned into a mammalian expression plasmid, the plasmid was transfected into human embryonic kidney 293T (HEK293T) cells. Thereafter, VmPyV VLPs were purified from the cell lysates of the transfected cells via sucrose gradient sedimentation. Electron microscopic analyses revealed that VmPyV VP1 forms VLPs with a diameter of approximately 50 nm that are exclusively localized in cell nuclei. Furthermore, we generated VLPs consisting of the deletion mutant VmPyV VP1 (ΔC VP1) lacking the C-terminal 116 amino acid residues and compared its VLP formation efficiency and morphology to those of VLPs from wild-type VmPyV VP1 (WT VP1). WT and ΔC VP1 VLPs were similar in size, but the number of ΔC VP1 VLPs was much lower than that of WT VP1 VLPs in VP1-expressing HEK293T cells. These results suggest that the length of VP1 is unrelated to virion morphology; however, the C-terminal region of VmPyV VP1 affects the efficiency of its VLP formation. PMID:24419975

  16. BK and JC polyomavirus infections in Tunisian renal transplant recipients.

    PubMed

    Boukoum, Hanen; Nahdi, Imen; Sahtout, Wissal; Skiri, Habib; Aloui, Sabra; Achour, Abdelatif; Segondy, Michel; Aouni, Mahjoub

    2015-10-01

    The aim of this prospective study was to investigate the rate of BK (BKPyV) and JC (JCPyV) polyomavirus infections and their influence on allograft function in Tunisian renal transplant recipients. A total of 72 renal transplant recipients were studied. BKPyV and JCPyV were detected and quantified by real-time PCR in urine and plasma. Demographic and laboratory characteristics were collected for each patient. Polyomavirus DNAuria was detected in 54 (75%) of renal transplant recipients: 26 (36%) had BKPyV DNAuria, 20 (28%) had JCPyV DNAuria, and 8 (11%) had a dual BKPyV/JCPyV DNAuria. BKPyV DNAemia was detected in four (5.5%) patients, whereas no patient had JCPyV viremia. More than 70% of BKPyV and JCPyV infections started within the first 3 months post-transplant. The risk for positive DNAemia was observed in patients with DNAuria level >10(7) copies/ml. BK Polyomavirus-associated nephropathy (BKPyVAN) was observed in two patients. This study highlights the high frequency of BKPyV and JCPyV viruria during the first year post-transplant with the highest incidence observed in the third month. We identified several risk factors that were associated with BKV DNAuria including age, sex of patients, and the use of tacrolimus instead of cyclosporine A at month 3. The use of cyclosporine A instead of tacrolimus was identified as risk factor for JCV viruria in month 3. No statistical difference in the allograft function was found between BKPyV and/or JCPyV infected and uninfected patients.

  17. Gene Regulation and Quality Control in Murine Polyomavirus Infection

    PubMed Central

    Carmichael, Gordon G.

    2016-01-01

    Murine polyomavirus (MPyV) infects mouse cells and is highly oncogenic in immunocompromised hosts and in other rodents. Its genome is a small, circular DNA molecule of just over 5000 base pairs and it encodes only seven polypeptides. While seemingly simply organized, this virus has adopted an unusual genome structure and some unusual uses of cellular quality control pathways that, together, allow an amazingly complex and varied pattern of gene regulation. In this review we discuss how MPyV leverages these various pathways to control its life cycle. PMID:27763514

  18. NASA Workshop on Technology for Human Robotic Exploration and Development of Space

    NASA Technical Reports Server (NTRS)

    Mankins, J. C.; Marzwell, N.; Mullins, C. A.; Christensen, C. B.; Howell, J. T.; O'Neil, D. A.

    2004-01-01

    Continued constrained budgets and growing interests in the industrialization and development of space requires NASA to seize every opportunity for assuring the maximum return on space infrastructure investments. This workshop provided an excellent forum for reviewing, evaluating, and updating pertinent strategic planning, identifying advanced concepts and high-risk/high-leverage research and technology requirements, developing strategies and roadmaps, and establishing approaches, methodologies, modeling, and tools for facilitating the commercial development of space and supporting diverse exploration and scientific missions. Also, the workshop addressed important topic areas including revolutionary space systems requiring investments in innovative advanced technologies; achieving transformational space operations through the insertion of new technologies; revolutionary science in space through advanced systems and new technologies enabling experiments to go anytime to any location; and, innovative and ambitious concepts and approaches essential for promoting advancements in space transportation. Details concerning the workshop process, structure, and results are contained in the ensuing report.

  19. Summary of the International Workshop of Dietary Sodium and Human Health in China and the United States.

    PubMed

    Alderman, Michael H

    2009-09-01

    The idea for a workshop focused on sodium and health was generated by the editors of the American Journal of Hypertension and the Chinese Journal of Hypertension as part of their emerging collaboration. It reflects a belief that scientists and clinicians interested in blood pressure and its related conditions would profit from more effective communication and interaction. Salt was chosen as a central topic because of its multiple physiological and pathological effects, its importance to human health, its role in elevating blood pressure, the unresolved scientific issues, and the absence of an agreed public health response regarding dietary sodium. In addition, much current clinical and population research has begun to shed light on how the interaction of genetics, environment, and diet determines the impact of sodium intake on human health and disease. Surprisingly, given the often passionate professional interest, there has been no regular forum dedicated to an exchange of new knowledge and current research findings. The biomedical literature provides strong evidence that investigators around the world are generating important new findings that increase understanding of how dietary sodium may relate to both health and disease. Given the widely shared need and interest, the workshop's organizers hope that the experience in Xi'an will lead to the appearance of regular working meetings on sodium and human disease on the international scientific schedule. The ultimate goal is for this scientific discourse to lead to evidence-based programs for population salt consumption that will contribute to the betterment of human health.

  20. Report of the Fourth International Workshop on human X chromosome mapping 1993

    SciTech Connect

    Schlessinger, D.; Mandel, J.L.; Monaco, A.P.; Nelson, D.L.; Willard, H.F.

    1993-12-31

    Vigorous interactive efforts by the X chromosome community have led to accelerated mapping in the last six months. Seventy-five participants from 12 countries around the globe contributed progress reports to the Fourth International X Chromosome Workshop, at St. Louis, MO, May 9-12, 1993. It became clear that well over half the chromosome is now covered by YAC contigs that are being extended, verified, and aligned by their content of STSs and other markers placed by cytogenetic or linkage mapping techniques. The major aim of the workshop was to assemble the consensus map that appears in this report, summarizing both consensus order and YAC contig information.

  1. 76 FR 50221 - International Workshop on Alternative Methods for Human and Veterinary Rabies Vaccine Testing...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-12

    ... testing, and to develop an implementation strategy to achieve global acceptance and use of these... necessary to achieve global acceptance and implementation. The workshop is organized by NICEATM, ICCVAM, the..., revised, and alternative test methods and strategies. NICEATM and ICCVAM welcome the public nomination...

  2. REPORT ON WORKSHOP ON SUSTAINABILITY AND INDUSTRY: ENERGY, MATERIAL CONSUMPTION, AND HUMAN BEHAVIOR

    EPA Science Inventory

    The purpose of the Workshop was to begin a process by which the leaders of the Council for Chemical Research, industry, academia, and government focus on sustainability and devote substantial resources to advancing issues that will improve the sustainability of industry and socie...

  3. Report of the fifth international workshop on human X chromosome mapping

    SciTech Connect

    Willard, H.F.; Cremers, F.; Mandel, J.L.; Monaco, A.P.; Nelson, D.L.; Schlessinger, D.

    1994-12-31

    A high-quality integrated genetic and physical map of the X chromosome from telomere to telomere, based primarily on YACs formatted with probes and STSs, is increasingly close to reality. At the Fifth International X Chromosome Workshop, organized by A.M. Poustka and D. Schlessinger in Heidelberg, Germany, April 24--27, 1994, substantial progress was recorded on extension and refinement of the physical map, on the integration of genetic and cytogenetic data, on attempts to use the map to direct gene searches, and on nascent large-scale sequencing efforts. This report summarizes physical and genetic mapping information presented at the workshop and/or published since the reports of the fourth International X Chromosome Workshop. The principle aim of the workshop was to derive a consensus map of the chromosome, in terms of physical contigs emphasizing the location of genes and microsatellite markers. The resulting map is presented and updates previous versions. This report also updates the list of highly informative microsatellites. The text highlights the working state of the map, the genes known to reside on the X, and the progress toward integration of various types of data.

  4. International Coordination of Large-Scale Human Induced Pluripotent Stem Cell Initiatives: Wellcome Trust and ISSCR Workshops White Paper

    PubMed Central

    Soares, Filipa A.C.; Sheldon, Michael; Rao, Mahendra; Mummery, Christine; Vallier, Ludovic

    2014-01-01

    There is growing recognition of the potential value of human induced pluripotent stem cells (hiPSC) for understanding disease and identifying drugs targets. This has been reflected in the establishment of multiple large-scale hiPSC initiatives worldwide. Representatives of these met recently at a workshop supported by the Welcome Trust in the UK and in a focus session at the 2014 ISSCR annual meeting in Vancouver. The purpose was to discuss strategies for making thousands of hiPSC lines widely available with as few restrictions as possible while retaining financial viability and donor privacy. The outcome of these discussions is described here. PMID:25496616

  5. Characterization of the DNA binding properties of polyomavirus capsid protein

    NASA Technical Reports Server (NTRS)

    Chang, D.; Cai, X.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    The DNA binding properties of the polyomavirus structural proteins VP1, VP2, and VP3 were studied by Southwestern analysis. The major viral structural protein VP1 and host-contributed histone proteins of polyomavirus virions were shown to exhibit DNA binding activity, but the minor capsid proteins VP2 and VP3 failed to bind DNA. The N-terminal first five amino acids (Ala-1 to Lys-5) were identified as the VP1 DNA binding domain by genetic and biochemical approaches. Wild-type VP1 expressed in Escherichia coli (RK1448) exhibited DNA binding activity, but the N-terminal truncated VP1 mutants (lacking Ala-1 to Lys-5 and Ala-1 to Cys-11) failed to bind DNA. The synthetic peptide (Ala-1 to Cys-11) was also shown to have an affinity for DNA binding. Site-directed mutagenesis of the VP1 gene showed that the point mutations at Pro-2, Lys-3, and Arg-4 on the VP1 molecule did not affect DNA binding properties but that the point mutation at Lys-5 drastically reduced DNA binding affinity. The N-terminal (Ala-1 to Lys-5) region of VP1 was found to be essential and specific for DNA binding, while the DNA appears to be non-sequence specific. The DNA binding domain and the nuclear localization signal are located in the same N-terminal region.

  6. Enhancer dependence of polyomavirus persistence in mouse kidneys.

    PubMed Central

    Rochford, R; Moreno, J P; Peake, M L; Villarreal, L P

    1992-01-01

    We previously showed that alterations in the enhancer sequence of polyomavirus DNA can alter both the level and the organ specificity of viral DNA replication during the acute phase of infection of newborn mice (R. Rochford, B. A. Campbell, and L. P. Villarreal, J. Virol. 64:476-485, 1990). In this study, we examined whether these enhancer sequence alterations can also affect polyomavirus replication during the persistent phase of infection in vivo. After infection of newborn mice with a mixture of three enhancer variants, the individual organs could select for enhancer-specific viral DNA replication during both the acute and the persistent phases of infection. Contrary to expectations, the ability of some variants to establish a high-level acute infection in some organs (e.g., the pancreas) did not necessarily lead to a persistent infection in those organs. Thus, enhancers can affect acute and persistent infections differently. In addition, some enhancer variants tended to establish a high-level persistent infection in the kidneys immediately following an acute infection; however, in all cases considerable histopathology was associated with these elevated long-term infections, and these mice were always runty. A persistent infection in the kidneys thus appears able to exist in two distinguishable states, a high-level pathological state and a low-level nonpathological state, which can be affected by the viral enhancer sequence. Images PMID:1316448

  7. Mapping of phosphorylation sites in polyomavirus large T antigen

    SciTech Connect

    Hassauer, M.; Scheidtmann, K.H.; Walter, G.

    1986-06-01

    The phosphorylation sites of polyomavirus large T antigen from infected or transformed cells were investigated. Tryptic digestion of large T antigen from infected, /sup 32/P/sub i/-labeled cells revealed seven major phosphopeptides. Five of these were phosphorylated only at serine residues, and two were phosphorylated at serine and threonine residues. The overall ratio of phosphoserine to phosphothreonine was 6:1. The transformed cell line B4 expressed two polyomavirus-specific phosphoproteins: large T antigen, which was only weakly phosphorylated, and a truncated form of large T antigen of 34,000 molecular weight which was heavily phosphorylated. Both showed phosphorylation patterns similar to that of large T antigen from infected cells. Peptide analyses of large T antigens encoded by the deletion mutants dl8 and dl23 or of specific fragments of wild-type large T antigen indicated that the phosphorylation sites are located in an amino-terminal region upstream of residue 194. The amino acid composition of the phosphopeptides as revealed by differential labeling with various amino acids indicated that several phosphopeptides contain overlapping sequences and that all phosphorylation sites are located in four tryptic peptides derived from a region between Met71 and Arg191. Two of the potential phosphorylation sites were identified as Ser81 and Thr187. The possible role of this modification of large T antigen is discussed.

  8. Building the Human Vaccines Project: strategic management recommendations and summary report of the 15-16 July 2014 business workshop.

    PubMed

    Schenkelberg, Theodore; Kieny, Marie-Paule; Bianco, A E; Koff, Wayne C

    2015-05-01

    The Human Vaccines Project is a bold new initiative, with the goal of solving the principal scientific problem impeding vaccine development for infectious diseases and cancers: the generation of specific, broad, potent and durable immune responses in humans. In the July 2014 workshop, 20 leaders from the public and private sectors came together to give input on strategic business issues for the creation of the Human Vaccines Project. Participants recommended the Project to be established as a nonprofit public-private partnership, structured as a global R&D consortium closely engaged with industrial partners, and located/affiliated with one or more major academic centers conducting vaccine R&D. If successful, participants concluded that the Project could greatly accelerate the development of new and improved vaccines, with the potential to transform disease prevention in the 21st century.

  9. CONCLUSIONS AND RECOMMENDATIONS OF THE EXPERT PANEL: TECHNICAL WORKSHOP ON HUMAN MILK SURVEILLANCE AND BIOMONITORING FOR ENVIRONMENTAL CHEMICALS IN THE UNITED STATES

    EPA Science Inventory

    The Technical Workshop focused on questions related to interpretation of information gathered from human milk biomonitoring studies. Biomonitoring can measure a person’s exposure to a chemical in his/her tissue. Human milk is a unique biological matrix for biomonitoring because i...

  10. Polyomavirus nephropathy of the native kidney in a patient with rheumatoid arthritis and pulmonary fibrosis.

    PubMed

    Krystel-Whittemore, Melissa; McCarthy, Ellen T; Damjanov, Ivan; Fields, Timothy A

    2015-08-28

    Polyomavirus nephropathy is commonly seen in the renal allograft setting but is uncommon in native kidneys. This paper describes polyomavirus nephropathy that developed in the native kidneys of a patient following immunosuppressive therapy for rheumatoid arthritis/Sjögren's syndrome associated lung disease. The patient presented with dyspnoea and a slow steady rise in serum creatinine. Owing to chronic immunosuppression, calcineurin-inhibitor toxicity was suspected. However, renal biopsy revealed polyomavirus nephropathy. The treatment of choice, lowered immunosuppression, was complicated by exacerbation of the patient's lung disease. This case highlights features of polyomavirus nephropathy in the native kidney, as well as the difficulty in its treatment when immunosuppressive treatment is necessary for medical comorbidities.

  11. Rapid detection of trichodysplasia spinulosa-associated polyomavirus in skin biopsy specimen.

    PubMed

    Urbano, Paulo Roberto P; Pannuti, Cláudio Sérgio; Pierrotti, Ligia C; David-Neto, Elias; Romano, Camila Malta

    2014-07-24

    Trichodysplasia spinulosa-associated polyomavirus (TSV) is responsible for a rare skin cancer. Using metagenomic approaches, we determined the complete genome sequence of a TSV first detected in Brazil in spicules of an immunocompromised patient suspected to have trichodysplasia spinulosa.

  12. High-Affinity Rb Binding, p53 Inhibition, Subcellular Localization, and Transformation by Wild-Type or Tumor-Derived Shortened Merkel Cell Polyomavirus Large T Antigens

    PubMed Central

    Borchert, Sophie; Czech-Sioli, Manja; Neumann, Friederike; Schmidt, Claudia; Wimmer, Peter; Dobner, Thomas

    2014-01-01

    ABSTRACT Interference with tumor suppressor pathways by polyomavirus-encoded tumor antigens (T-Ags) can result in transformation. Consequently, it is thought that T-Ags encoded by Merkel cell polyomavirus (MCPyV), a virus integrated in ∼90% of all Merkel cell carcinoma (MCC) cases, are major contributors to tumorigenesis. The MCPyV large T-Ag (LT-Ag) has preserved the key functional domains present in all family members but has also acquired unique regions that flank the LxCxE motif. As these regions may mediate unique functions, or may modulate those shared with T-Ags of other polyomaviruses, functional studies of MCPyV T-Ags are required. Here, we have performed a comparative study of full-length or MCC-derived truncated LT-Ags with regard to their biochemical characteristics, their ability to bind to retinoblastoma (Rb) and p53 proteins, and their transforming potential. We provide evidence that full-length MCPyV LT-Ag may not directly bind to p53 but nevertheless can significantly reduce p53-dependent transcription in reporter assays. Although early region expression constructs harboring either full-length or MCC-derived truncated LT-Ag genes can transform primary baby rat kidney cells, truncated LT-Ags do not bind to p53 or reduce p53-dependent transcription. Interestingly, shortened LT-Ags exhibit a very high binding affinity for Rb, as shown by coimmunoprecipitation and in vitro binding studies. Additionally, we show that truncated MCPyV LT-Ag proteins are expressed at higher levels than those for the wild-type protein and are able to partially relocalize Rb to the cytoplasm, indicating that truncated LT proteins may have gained additional features that distinguish them from the full-length protein. IMPORTANCE MCPyV is one of the 12 known polyomaviruses that naturally infect humans. Among these, it is of particular interest since it is the only human polyomavirus known to be involved in tumorigenesis. MCPyV is thought to be causally linked to MCC, a rare

  13. Progressive multifocal leukoencephalopathy-associated mutations in the JC polyomavirus capsid disrupt lactoseries tetrasaccharide c binding.

    PubMed

    Maginnis, Melissa S; Ströh, Luisa J; Gee, Gretchen V; O'Hara, Bethany A; Derdowski, Aaron; Stehle, Thilo; Atwood, Walter J

    2013-06-11

    The human JC polyomavirus (JCPyV) is the causative agent of the fatal, demyelinating disease progressive multifocal leukoencephalopathy (PML). The Mad-1 prototype strain of JCPyV uses the glycan lactoseries tetrasaccharide c (LSTc) and serotonin receptor 5-HT2A to attach to and enter into host cells, respectively. Specific residues in the viral capsid protein VP1 are responsible for direct interactions with the α2,6-linked sialic acid of LSTc. Viral isolates from individuals with PML often contain mutations in the sialic acid-binding pocket of VP1 that are hypothesized to arise from positive selection. We reconstituted these mutations in the Mad-1 strain of JCPyV and found that they were not capable of growth. The mutations were then introduced into recombinant VP1 and reconstituted as pentamers in order to conduct binding studies and structural analyses. VP1 pentamers carrying PML-associated mutations were not capable of binding to permissive cells. High-resolution structure determination revealed that these pentamers are well folded but no longer bind to LSTc due to steric clashes in the sialic acid-binding site. Reconstitution of the mutations into JCPyV pseudoviruses allowed us to directly quantify the infectivity of the mutants in several cell lines. The JCPyV pseudoviruses with PML-associated mutations were not infectious, nor were they able to engage sialic acid as measured by hemagglutination of human red blood cells. These results demonstrate that viruses from PML patients with single point mutations in VP1 disrupt binding to sialic acid motifs and render these viruses noninfectious. IMPORTANCE Infection with human JC polyomavirus (JCPyV) is common and asymptomatic in healthy individuals, but during immunosuppression, JCPyV can spread from the kidney to the central nervous system (CNS) and cause a fatal, demyelinating disease, progressive multifocal leukoencephalopathy (PML). Individuals infected with HIV, those who have AIDS, or those receiving

  14. Report on the international workshop on alternative methods for human and veterinary rabies vaccine testing: state of the science and planning the way forward.

    PubMed

    Stokes, William; McFarland, Richard; Kulpa-Eddy, Jodie; Gatewood, Donna; Levis, Robin; Halder, Marlies; Pulle, Gayle; Kojima, Hajime; Casey, Warren; Gaydamaka, Alexander; Miller, Timothy; Brown, Karen; Lewis, Charles; Chapsal, Jean-Michel; Bruckner, Lukas; Gairola, Sunil; Kamphuis, Elisabeth; Rupprecht, Charles E; Wunderli, Peter; McElhinney, Lorraine; De Mattia, Fabrizio; Gamoh, Koichiro; Hill, Richard; Reed, David; Doelling, Vivian; Johnson, Nelson; Allen, David; Rinckel, Lori; Jones, Brett

    2012-09-01

    Potency testing of most human and veterinary rabies vaccines requires vaccination of mice followed by a challenge test using an intracerebral injection of live rabies virus. NICEATM, ICCVAM, and their international partners organized a workshop to review the availability and validation status of alternative methods that might reduce, refine, or replace the use of animals for rabies vaccine potency testing, and to identify research and development efforts to further advance alternative methods. Workshop participants agreed that general anesthesia should be used for intracerebral virus injections and that humane endpoints should be used routinely as the basis for euthanizing animals when conducting the mouse rabies challenge test. Workshop participants recommended as a near-term priority replacement of the mouse challenge with a test validated to ensure potency, such as the mouse antibody serum neutralization test for adjuvanted veterinary rabies vaccines for which an international collaborative study was recently completed. The workshop recommended that an in vitro antigen quantification test should be a high priority for product-specific validation of human and non-adjuvanted veterinary rabies vaccines. Finally, workshop participants recommended greater international cooperation to expedite development, validation, regulatory acceptance, and implementation of alternative test methods for rabies vaccine potency testing.

  15. Workshop Review

    ERIC Educational Resources Information Center

    Journal of Aerospace Education, 1977

    1977-01-01

    Reviews a leadership development aerospace educators workshop held at Maxwell Air Force Base, Alabama, July 22, 1977, and an introductory/advanced aerospace workshop held at Central Washington State College. (SL)

  16. Characterization of California sea lion polyomavirus 1: expansion of the known host range of the Polyomaviridae to Carnivora.

    PubMed

    Wellehan, James F X; Rivera, Rebecca; Archer, Linda L; Benham, Celeste; Muller, Jennifer K; Colegrove, Kathleen M; Gulland, Frances M D; St Leger, Judy A; Venn-Watson, Stephanie K; Nollens, Hendrik H

    2011-07-01

    The genome of a novel polyomavirus first identified in a proliferative tongue lesion of a California sea lion (Zalophus californianus) is reported. This is only the third described polyomavirus of laurasiatherian mammals, is the first of the three associated with a lesion, and is the first known polyomavirus of a host in the order Carnivora. Predicted large T, small t, VP1, VP2, and VP3 genes were identified based on homology to proteins of known polyomaviruses, and a putative agnoprotein was identified based upon its location in the genome. Phylogenetic analysis of the predicted late region proteins found that the laurasiatherian polyomaviruses, together with Squirrel monkey polyomavirus and Murine pneumotropic virus, form a monophyletic clade. Phylogenetic analysis of the early region was more ambiguous. The noncoding control region of California sea lion polyomavirus 1 is unusual in that only two apparent large T binding sites are present; this is less than any other known polyomavirus. The VP1 of this virus has an unusually long carboxy-terminal region. A quantitative polymerase chain reaction was developed and utilized on various samples from 79 additional animals from either managed or wild stranded California sea lion populations, and California sea lion polyomavirus 1 infection was found in 24% of stranded animals. Sequence of additional samples identified four sites of variation in the t antigens, three of which resulted in predicted coding changes.

  17. BK Polyomavirus: Clinical Aspects, Immune Regulation, and Emerging Therapies.

    PubMed

    Ambalathingal, George R; Francis, Ross S; Smyth, Mark J; Smith, Corey; Khanna, Rajiv

    2017-04-01

    BK polyomavirus (BKV) causes frequent infections during childhood and establishes persistent infections within renal tubular cells and the uroepithelium, with minimal clinical implications. However, reactivation of BKV in immunocompromised individuals following renal or hematopoietic stem cell transplantation may cause serious complications, including BKV-associated nephropathy (BKVAN), ureteric stenosis, or hemorrhagic cystitis. Implementation of more potent immunosuppression and increased posttransplant surveillance has resulted in a higher incidence of BKVAN. Antiviral immunity plays a crucial role in controlling BKV replication, and our increasing knowledge about host-virus interactions has led to the development of improved diagnostic tools and clinical management strategies. Currently, there are no effective antiviral agents for BKV infection, and the mainstay of managing reactivation is reduction of immunosuppression. Development of immune-based therapies to combat BKV may provide new and exciting opportunities for the successful treatment of BKV-associated complications.

  18. JC Polyomavirus (JCV) and Monoclonal Antibodies: Friends or Potential Foes?

    PubMed Central

    Clementi, Nicola; Mancini, Nicasio; Solforosi, Laura; Clementi, Massimo

    2013-01-01

    Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system (CNS), observed in immunodeficient patients and caused by JC virus ((JCV), also called JC polyomavirus (JCPyV)). After the HIV pandemic and the introduction of immunomodulatory therapy, the PML incidence significantly increased. The correlation between the use of natalizumab, a drug used in multiple sclerosis (MS), and the PML development of particular relevance. The high incidence of PML in natalizumab-treated patients has highlighted the importance of two factors: the need of PML risk stratification among natalizumab-treated patients and the need of effective therapeutic options. In this review, we discuss these two needs under the light of the major viral models of PML etiopathogenesis. PMID:23878587

  19. Interactions among the major and minor coat proteins of polyomavirus.

    PubMed Central

    Barouch, D H; Harrison, S C

    1994-01-01

    Murine polyomavirus contains two related minor coat proteins, VP2 and VP3, in addition to the major coat protein, VP1. The sequence of VP3 is identical to that of the carboxy-terminal two-thirds of VP2. VP2 may serve a role in uncoating of the virus, and both minor coat proteins may be important for viral assembly. In this study, we show that VP3 and a series of deletion mutants of VP3 can be expressed in Escherichia coli as fusion proteins to glutathione S-transferase and partially solubilized with a mild detergent. Using an in vitro binding assay, we demonstrate that a 42-amino-acid fragment near the carboxy terminus of VP3 (residues 140 to 181) is sufficient for binding to purified VP1 pentamers. This binding interaction is rapid, saturable, and specific for the common carboxy terminus of VP2 and VP3. The VP1-VP3 complex can be coimmunoprecipitated with an antibody specific to VP1, and a purified VP3 fragment can selectively extract VP1 from a crude cell lysate. The stoichiometry of the binding reaction suggests that each VP1 pentamer in the virus binds either one VP2 or one VP3, with the VP1-VP2/3 complex stabilized by hydrophobic interactions. These results, taken together with studies from other laboratories on the expression of polyomavirus capsid proteins in mouse and insect cells (S. E. Delos, L. Montross, R. B. Moreland, and R. L. Garcea, Virology, 194:393-398, 1993; J. Forstova, N. Krauzewicz, S. Wallace, A. J. Street, S. M. Dilworth, S. Beard, and B. E. Griffin, J. Virol. 67:1405-1413, 1993), support the idea that a VP1-VP2/3 complex forms in the cytoplasm and, after translocation into the nucleus, acts as the unit for viral assembly. Images PMID:8189532

  20. DOE Human Genome Program: Contractor-Grantee Workshop IV, November 13--17, 1994, Santa Fe, New Mexico

    SciTech Connect

    Not Available

    1994-10-01

    This volume contains the proceedings of the fourth Contractor-Grantee Workshop for the Department of Energy (DOE) Human Genome Program. Of the 204 abstracts in this book, some 200 describe the genome research of DOE-funded grantees and contractors located at the multidisciplinary centers at Lawrence Berkeley Laboratory, Lawrence Livermore National Laboratory, and Los Alamos National Laboratory; other DOE-supported laboratories; and more than 54 universities, research organizations, and companies in the United States and abroad. Included are 16 abstracts from ongoing projects in the Ethical, Legal, and Social Issues (ELSI) component, an area that continues to attract considerable attention from a wide variety of interested parties. Three abstracts summarize work in the new Microbial Genome Initiative launched this year by the Office of Health and Environmental Research (OHER) to provide genome sequence and mapping data on industrially important microorganisms and those that live under extreme conditions. Many of the projects will be discussed at plenary sessions held throughout the workshop, and all are represented in the poster sessions.

  1. Commercially available immunoglobulins contain virus neutralizing antibodies against all major genotypes of polyomavirus BK.

    PubMed

    Randhawa, P; Pastrana, D V; Zeng, G; Huang, Y; Shapiro, R; Sood, P; Puttarajappa, C; Berger, M; Hariharan, S; Buck, C B

    2015-04-01

    Neutralizing antibodies (NAbs) form the basis of immunotherapeutic strategies against many important human viral infections. Accordingly, we studied the prevalence, titer, genotype-specificity, and mechanism of action of anti-polyomavirus BK (BKV) NAbs in commercially available human immune globulin (IG) preparations designed for intravenous (IV) use. Pseudovirions (PsV) of genotypes Ia, Ib2, Ic, II, III, and IV were generated by co-transfecting a reporter plasmid encoding luciferase and expression plasmids containing synthetic codon-modified VP1, VP2, and VP3 capsid protein genes into 293TT cells. NAbs were measured using luminometry. All IG preparations neutralized all BKV genotypes, with mean EC50 titers as high as 254 899 for genotype Ia and 6,666 for genotype IV. Neutralizing titers against genotypes II and III were higher than expected, adding to growing evidence that infections with these genotypes are more common than currently appreciated. Batch to batch variation in different lots of IG was within the limits of experimental error. Antibody mediated virus neutralizing was dose dependent, modestly enhanced by complement, genotype-specific, and achieved without effect on viral aggregation, capsid morphology, elution, or host cell release. IG contains potent NAbs capable of neutralizing all major BKV genotypes. Clinical trials based on sound pharmacokinetic principles are needed to explore prophylactic and therapeutic applications of these anti-viral effects, until effective small molecule inhibitors of BKV replication can be developed.

  2. An official American Thoracic Society workshop report: Climate change and human health.

    PubMed

    Pinkerton, Kent E; Rom, William N; Akpinar-Elci, Muge; Balmes, John R; Bayram, Hasan; Brandli, Otto; Hollingsworth, John W; Kinney, Patrick L; Margolis, Helene G; Martin, William J; Sasser, Erika N; Smith, Kirk R; Takaro, Tim K

    2012-03-01

    This document presents the proceedings from the American Thoracic Society Climate Change and Respiratory Health Workshop that was held on May 15, 2010, in New Orleans, Louisiana. The purpose of the one-day meeting was to address the threat to global respiratory health posed by climate change. Domestic and international experts as well as representatives of international respiratory societies and key U.S. federal agencies convened to identify necessary research questions concerning climate change and respiratory health and appropriate mechanisms and infrastructure needs for answering these questions. After much discussion, a breakout group compiled 27 recommendations for physicians, researchers, and policy makers. These recommendations are listed under main issues that the workshop participants deemed of key importance to respiratory health. Issues include the following: (1) the health impacts of climate change, with specific focus on the effect of heat waves, air pollution, and natural cycles; (2) mitigation and adaptation measures to be taken, with special emphasis on recommendations for the clinical and research community; (3) recognition of challenges specific to low-resource countries when coping with respiratory health and climate change; and (4) priority research infrastructure needs, with special discussion of international needs for cooperating with present and future environmental monitoring and alert systems.

  3. Dialogue with Asia's Rural Man. A Report of the Development of Human Resources in Rural Asia Workshop (DHRRAW) (Swanganivas, Thailand, August 4-25, 1974).

    ERIC Educational Resources Information Center

    Ledesma, Antonio L., Comp.; And Others

    General objective of the workshop was to provide a forum for Asian rural leaders in which they could discern the significance and implication of crucial processes by which human potentialities are harnessed for the integral growth of rural peoples and communities. The central question addressed was how to combine economic growth with social…

  4. Role of the Human Polyomavirus, BKV, in Prostate Cancer

    DTIC Science & Technology

    2005-08-01

    and late regions, and encodes five major proteins nephropathy (Arthur et al., 1986; Hiraoka et al., 1991; (Figure 1; Seif et al., 1979; Frisque et al...hematoxylin and eosin to confirm diagnosis . The The program used for fl-globin amplification consisted of samples were a mixture of three different...transplantation, pregnancy , or HIV infection, high throughout the rest of the time course, levels of BKV replication are observed in the kidneys and The next step

  5. Merkel cell polyomavirus infection in both components of a combined Merkel cell carcinoma and basal cell carcinoma with ductal differentiation; each component had a similar but different novel Merkel cell polyomavirus large T antigen truncating mutation.

    PubMed

    Iwasaki, Takeshi; Kodama, Hajime; Matsushita, Michiko; Kuroda, Naoto; Yamasaki, Yoshikazu; Murakami, Ichiro; Yamamoto, Osamu; Hayashi, Kazuhiko

    2013-03-01

    Merkel cell polyomavirus infects up to 80% of patients with Merkel cell carcinoma. Combined Merkel cell carcinoma and cutaneous tumors occur occasionally. Previous reports have suggested that Merkel cell polyomavirus is absent from combined Merkel cell carcinoma and squamous cell carcinomas. This is the first report that Merkel cell polyomavirus infected in both lesions of a combined Merkel cell carcinoma and basal cell carcinoma. A 92-year-old Japanese man presented with a right thigh small subcutaneous mass. Histologic examination revealed a combined tumor with Merkel cell carcinoma and basal cell carcinoma with ductal differentiation. Both tumors and intermingled Merkel cells in basal cell carcinoma expressed Merkel cell polyomavirus large T antigen, and 17 and 240 copies of Merkel cell polyomavirus/cell were detected in the microdissected Merkel cell carcinoma and basal cell carcinoma specimens, respectively. Mutation analysis of Merkel cell polyomavirus large T antigen revealed a novel truncating mutation in Merkel cell carcinoma and a similar but different mutation in the basal cell carcinoma. These results suggest that each was infected by a different Merkel cell polyomavirus subclone derived from a single Merkel cell polyomavirus.

  6. An analysis of myeloma plasma cell phenotype using antibodies defined at the IIIrd International Workshop on Human Leucocyte Differentiation Antigens.

    PubMed Central

    Jackson, N; Ling, N R; Ball, J; Bromidge, E; Nathan, P D; Franklin, I M

    1988-01-01

    Fresh bone marrow from 43 cases of myeloma and three cases of plasma cell leukaemia has been phenotyped both by indirect immune-rosetting and, on fixed cytospin preparations, by indirect immunofluorescence. Both clustered and unclustered B cell associated antibodies from the IIIrd International Workshop on Human Leucocyte Differentiation Antigens were used. The results confirm the lack of many pan-B antigens on the surface of myeloma plasma cells, i.e. CD19-23, 37, 39, w40. Strong surface reactivity is seen with CD38 antibodies and with one CD24 antibody (HB8). Weak reactions are sometimes obtained with CD9, 10 and 45R. On cytospin preparations CD37, 39 and w40 are sometimes weakly positive, and anti-rough endoplasmic reticulum antibodies are always strongly positive. Specific and surface-reacting antiplasma cell antibodies are still lacking. PMID:3048803

  7. Genome Sequences of Polyomaviruses from the Wild-Living Red Colobus (Piliocolobus badius) and Western Chimpanzee (Pan troglodytes verus).

    PubMed

    Ben Salem, Nicole; Leendertz, Fabian H; Ehlers, Bernhard

    2016-10-13

    We identified with PCR and sequencing the full genomes of the recently discovered Pan troglodytes verus polyomavirus 8 and Piliocolobus badius polyomavirus 2 in a western chimpanzee and a western red colobus free-ranging in Taï National Park of Côte d'Ivoire.

  8. Tumorigenic activity of Merkel cell polyomavirus T antigens expressed in the stratified epithelium of mice

    PubMed Central

    Spurgeon, Megan E.; Cheng, Jingwei; Bronson, Roderick T.; Lambert, Paul F.; DeCaprio, James A.

    2015-01-01

    Merkel cell polyomavirus (MCPyV) is frequently associated with Merkel cell carcinoma (MCC), a highly aggressive neuroendocrine skin cancer. Most MCC tumors contain integrated copies of the viral genome with persistent expression of the MCPyV large T (LT) and small T (ST) antigen. MCPyV isolated from MCC typically contain wild type ST but truncated forms of LT that retain the N-terminus but delete the C-terminus and render LT incapable of supporting virus replication. To determine the oncogenic activity of MCC tumor-derived T antigens in vivo, a conditional, tissue-specific mouse model was developed. Keratin 14-mediated Cre recombinase expression induced expression of MCPyV T antigens in stratified squamous epithelial cells and Merkel cells of the skin epidermis. Mice expressing MCPyV T antigens developed hyperplasia, hyperkeratosis, and acanthosis of the skin with additional abnormalities in whisker pads, footpads and eyes. Nearly half of the mice also developed cutaneous papillomas. Evidence for neoplastic progression within stratified epithelia included increased cellular proliferation, unscheduled DNA synthesis, increased E2F-responsive genes levels, disrupted differentiation, and presence of a DNA damage response. These results indicate that MCPyV T antigens are tumorigenic in vivo, consistent with their suspected etiological role in human cancer. PMID:25596282

  9. Merkel Cell Polyomavirus Small T Antigen Promotes Pro-Glycolytic Metabolic Perturbations Required for Transformation

    PubMed Central

    Keibler, Mark A.; Park, Donglim Esther; Molla, Vadim; Cheng, Jingwei; Stephanopoulos, Gregory

    2016-01-01

    Merkel cell polyomavirus (MCPyV) is an etiological agent of Merkel cell carcinoma (MCC), a highly aggressive skin cancer. The MCPyV small tumor antigen (ST) is required for maintenance of MCC and can transform normal cells. To gain insight into cellular perturbations induced by MCPyV ST, we performed transcriptome analysis of normal human fibroblasts with inducible expression of ST. MCPyV ST dynamically alters the cellular transcriptome with increased levels of glycolytic genes, including the monocarboxylate lactate transporter SLC16A1 (MCT1). Extracellular flux analysis revealed increased lactate export reflecting elevated aerobic glycolysis in ST expressing cells. Inhibition of MCT1 activity suppressed the growth of MCC cell lines and impaired MCPyV-dependent transformation of IMR90 cells. Both NF-κB and MYC have been shown to regulate MCT1 expression. While MYC was required for MCT1 induction, MCPyV-induced MCT1 levels decreased following knockdown of the NF-κB subunit RelA, supporting a synergistic activity between MCPyV and MYC in regulating MCT1 levels. Several MCC lines had high levels of MYCL and MYCN but not MYC. Increased levels of MYCL was more effective than MYC or MYCN in increasing extracellular acidification in MCC cells. Our results demonstrate the effects of MCPyV ST on the cellular transcriptome and reveal that transformation is dependent, at least in part, on elevated aerobic glycolysis. PMID:27880818

  10. Does polyomavirus infection interfere with bladder cancer fluorescence in situ hybridization?

    PubMed

    Hossain, Deloar; Hull, David; Kalantarpour, Fatemeh; Maitlen, Rebecca; Qian, Junqi; Bostwick, David G

    2014-03-01

    Urine cytology is a proven and widely used screening tool for the detection of urothelial carcinoma. However, morphologic features of polyomavirus infected cells, characterized by nuclear inclusions (decoy cells) are a known source of diagnostic confusion with malignancy. Fluorescence in situ hybridization (FISH) is now routinely used to support the cytological diagnosis of urothelial carcinoma and monitor for recurrence. We sought to determine whether polyomavirus infection could result in positive FISH results (aneuploidy). This study deals with retrospective study of 100 polyomavirus-infected urine samples from patients with no history of urothelial carcinoma or organ transplantation. All cases were stained with Papanicolaou and acid hematoxylin stain. One slide from each sample was de-stained and FISH was performed using chromosome enumeration probes 3, 7, 17, and locus-specific probe 9p21. Adequate cells for FISH analysis (25 cells) were present in 81 cases; 19 cases were insufficient due to loss of cells during de-staining and FISH preparation process. All polyomavirus-infected cells (decoy cells) exhibited a normal chromosome pattern. Four cases were FISH positive, but there were no positive decoy cells. Decoy cells did not exhibit aneuploidy by FISH. The presence of decoy cells does not exclude the possibility of concurrent urothelial carcinoma. Acid hematoxylin stain appeared to supplement the Papanicolou stain in identifying and confirming the presence of polyomavirus infection.

  11. Modulation of a Pore in the Capsid of JC Polyomavirus Reduces Infectivity and Prevents Exposure of the Minor Capsid Proteins

    PubMed Central

    Nelson, Christian D. S.; Ströh, Luisa J.; Gee, Gretchen V.; O'Hara, Bethany A.; Stehle, Thilo

    2015-01-01

    ABSTRACT JC polyomavirus (JCPyV) infection of immunocompromised individuals results in the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML). The viral capsid of JCPyV is composed primarily of the major capsid protein virus protein 1 (VP1), and pentameric arrangement of VP1 monomers results in the formation of a pore at the 5-fold axis of symmetry. While the presence of this pore is conserved among polyomaviruses, its functional role in infection or assembly is unknown. Here, we investigate the role of the 5-fold pore in assembly and infection of JCPyV by generating a panel of mutant viruses containing amino acid substitutions of the residues lining this pore. Multicycle growth assays demonstrated that the fitness of all mutants was reduced compared to that of the wild-type virus. Bacterial expression of VP1 pentamers containing substitutions to residues lining the 5-fold pore did not affect pentamer assembly or prevent association with the VP2 minor capsid protein. The X-ray crystal structures of selected pore mutants contained subtle changes to the 5-fold pore, and no other changes to VP1 were observed. Pore mutant pseudoviruses were not deficient in assembly, packaging of the minor capsid proteins, or binding to cells or in transport to the host cell endoplasmic reticulum. Instead, these mutant viruses were unable to expose VP2 upon arrival to the endoplasmic reticulum, a step that is critical for infection. This study demonstrated that the 5-fold pore is an important structural feature of JCPyV and that minor modifications to this structure have significant impacts on infectious entry. IMPORTANCE JCPyV is an important human pathogen that causes a severe neurological disease in immunocompromised individuals. While the high-resolution X-ray structure of the major capsid protein of JCPyV has been solved, the importance of a major structural feature of the capsid, the 5-fold pore, remains poorly understood. This pore is conserved across

  12. Human genetic research, race, ethnicity and the labeling of populations: recommendations based on an interdisciplinary workshop in Japan

    PubMed Central

    2014-01-01

    Background A challenge in human genome research is how to describe the populations being studied. The use of improper and/or imprecise terms has the potential to both generate and reinforce prejudices and to diminish the clinical value of the research. The issue of population descriptors has not attracted enough academic attention outside North America and Europe. In January 2012, we held a two-day workshop, the first of its kind in Japan, to engage in interdisciplinary dialogue between scholars in the humanities, social sciences, medical sciences, and genetics to begin an ongoing discussion of the social and ethical issues associated with population descriptors. Discussion Through the interdisciplinary dialogue, we confirmed that the issue of race, ethnicity and genetic research has not been extensively discussed in certain Asian communities and other regions. We have found, for example, the continued use of the problematic term, “Mongoloid” or continental terms such as “European,” “African,” and “Asian,” as population descriptors in genetic studies. We, therefore, introduce guidelines for reporting human genetic studies aimed at scientists and researchers in these regions. Conclusion We need to anticipate the various potential social and ethical problems entailed in population descriptors. Scientists have a social responsibility to convey their research findings outside of their communities as accurately as possible, and to consider how the public may perceive and respond to the descriptors that appear in research papers and media articles. PMID:24758583

  13. n-Butyrate, a cell cycle blocker, inhibits the replication of polyomaviruses and papillomaviruses but not that of adenoviruses and herpesviruses.

    PubMed Central

    Shadan, F F; Cowsert, L M; Villarreal, L P

    1994-01-01

    Small DNA viruses are dependent on the interaction of early proteins (such as large T antigen) with host p53 and Rb to bring about the G1-to-S cell cycle transition. The large DNA viruses are less dependent on host regulatory genes since additional early viral proteins (such as viral DNA polymerase, DNA metabolic enzymes, and other replication proteins) are involved in DNA synthesis. A highly conserved domain of large T antigen (similar to the p53-binding region) exclusively identifies papovavirus, parvovirus, and papillomaviruses from all other larger DNA viruses and implies a conserved interaction with host regulatory genes. In this report, we show that 3 to 6 mM butyrate, a general cell cycle blocker implicated in inhibition of the G1-to-S transition, inhibits DNA replication of polyomavirus and human papillomavirus type 11 but not the replication of larger DNA viruses such as adenovirus types 2 and 5, herpes simplex virus type 1, Epstein-Barr virus, and cytomegalovirus, which all bypass the butyrate-mediated cell cycle block. This butyrate effect on polyomavirus replication is not cell type specific, nor does it depend on the p53 or Rb gene, as inhibition was seen in fibroblasts with intact or homozygous deleted p53 or Rb, 3T6 cells, keratinocytes, C2C12 myoblasts, and 3T3-L1 adipocytes. In addition, butyrate did not inhibit expression of polyomavirus T antigen. The antiviral effect of butyrate involves a form of imprinted state, since pretreatment of cells with 3 mM butyrate inhibits human papillomavirus type 11 DNA replication for at least 96 h after its removal. Butyrate, therefore, serves as a molecular tool in dissecting the life cycle of smaller DNA viruses from that of the larger DNA viruses in relation to the cell cycle. Images PMID:8035479

  14. Nuclear localization of Merkel cell polyomavirus large T antigen in Merkel cell carcinoma

    SciTech Connect

    Nakamura, Tomoyuki; Sato, Yuko; Watanabe, Daisuke; Ito, Hideki; Shimonohara, Nozomi; Tsuji, Takahiro; Nakajima, Noriko; Suzuki, Yoshio; Matsuo, Koma; Nakagawa, Hidemi; Sata, Tetsutaro; Katano, Harutaka

    2010-03-15

    To clarify whether mutations in the large T gene encoded by Merkel cell polyomavirus affect the expression and function of large T antigen in Merkel cell carcinoma cases, we investigated the expression of large T antigen in vitro and in vivo. Immunohistochemistry using a rabbit polyclonal antibody revealed that large T antigen was expressed in the nuclei of Merkel cell carcinoma cells with Merkel cell polyomavirus infection. Deletion mutant analyses identified an Arg-Lys-Arg-Lys sequence (amino acids 277-280) as a nuclear localization signal in large T antigen. Sequence analyses revealed that there were no mutations in the nuclear localization signal in any of the eleven Merkel cell polyomavirus strains examined. Furthermore, stop codons were not observed in the upstream of the nuclear localization signal in any of the Merkel cell carcinoma cases examined. These data suggest that the nuclear localization signal is highly conserved and functional in Merkel cell carcinoma cases.

  15. Identification of a Novel Cetacean Polyomavirus from a Common Dolphin (Delphinus delphis) with Tracheobronchitis

    PubMed Central

    Anthony, Simon J.; St. Leger, Judy A.; Navarrete-Macias, Isamara; Nilson, Erica; Sanchez-Leon, Maria; Liang, Eliza; Seimon, Tracie; Jain, Komal; Karesh, William; Daszak, Peter; Briese, Thomas; Lipkin, W. Ian

    2013-01-01

    A female short-beaked common dolphin calf was found stranded in San Diego, California in October 2010, presenting with multifocal ulcerative lesions in the trachea and bronchi. Viral particles suggestive of polyomavirus were detected by EM, and subsequently confirmed by PCR and sequencing. Full genome sequencing (Ion Torrent) revealed a circular dsDNA genome of 5,159 bp that was shown to form a distinct lineage within the genus Polyomavirus based on phylogenetic analysis of the early and late transcriptomes. Viral infection and distribution in laryngeal mucosa was characterised using in-situ hybridisation, and apoptosis observed in the virus-infected region. These results demonstrate that polyomaviruses can be associated with respiratory disease in cetaceans, and expand our knowledge of their diversity and clinical significance in marine mammals. PMID:23874559

  16. Identification of a novel cetacean polyomavirus from a common dolphin (Delphinus delphis) with Tracheobronchitis.

    PubMed

    Anthony, Simon J; St Leger, Judy A; Navarrete-Macias, Isamara; Nilson, Erica; Sanchez-Leon, Maria; Liang, Eliza; Seimon, Tracie; Jain, Komal; Karesh, William; Daszak, Peter; Briese, Thomas; Lipkin, W Ian

    2013-01-01

    A female short-beaked common dolphin calf was found stranded in San Diego, California in October 2010, presenting with multifocal ulcerative lesions in the trachea and bronchi. Viral particles suggestive of polyomavirus were detected by EM, and subsequently confirmed by PCR and sequencing. Full genome sequencing (Ion Torrent) revealed a circular dsDNA genome of 5,159 bp that was shown to form a distinct lineage within the genus Polyomavirus based on phylogenetic analysis of the early and late transcriptomes. Viral infection and distribution in laryngeal mucosa was characterised using in-situ hybridisation, and apoptosis observed in the virus-infected region. These results demonstrate that polyomaviruses can be associated with respiratory disease in cetaceans, and expand our knowledge of their diversity and clinical significance in marine mammals.

  17. Simultaneous BK Polyomavirus (BKPyV)-associated nephropathy and hemorrhagic cystitis after living donor kidney transplantation.

    PubMed

    Helanterä, Ilkka; Hirsch, Hans H; Wernli, Marion; Ortiz, Fernanda; Lempinen, Marko; Räisänen-Sokolowski, Anne; Auvinen, Eeva; Mannonen, Laura; Lautenschlager, Irmeli

    2016-03-01

    BK polyomavirus (BKPyV) commonly reactivates after kidney transplantation, and can cause polyomavirus-associated nephropathy (PyVAN), whereas after allogeneic stem cell transplantation the most frequent manifestation of BKPyV is polyomavirus-associated hemorrhagic cystitis (PyVHC). Despite high-level BKPyV replication in both, the pathogenesis and manifestation of both BKPyV entities appears to differ substantially. We describe an unusual case of simultaneous PyVAN and PyVHC presenting with acute symptoms in a BKPyV-IgG positive recipient eight months after kidney transplantation from a haploidentical living donor, who was BKPyV-IgG negative. Symptoms of cystitis and viremia subsided rapidly after reduction of immunosuppression.

  18. The KIND Workshop Leader's Guide.

    ERIC Educational Resources Information Center

    Sirch, Willow Ann

    The National Association for Humane and Environmental Education (NAHEE) produces this workshop guide which offers a scripted workshop with handouts for elementary educators interested in sharing curriculum-based activities dedicated to helping young people develop values of kindness and respect toward people, animals, and the Earth. This guide…

  19. Strengthening the Paediatricians Project 2: The effectiveness of a workshop to address the Priority Mental Health Disorders of adolescence in low-health related human resource countries

    PubMed Central

    2010-01-01

    Background Paediatricians can be empowered to address the Priority Mental Health Disorders at primary care level. To evaluate the effectiveness of a collaborative workshop in enhancing the adolescent psychiatry knowledge among paediatricians. Methods A 3-day, 27-hours workshop was held for paediatricians from different regions of India under the auspices of the National Adolescent Paediatric Task Force of the Indian Academy of Paediatrics. A 5-item pretest-posttest questionnaire was developed and administered at the beginning and end of the workshop to evaluate the participants' knowledge acquisition in adolescent psychiatry. Bivariate and multivariate analyses were performed on an intention-to-participate basis. Results Forty-eight paediatricians completed the questionnaire. There was significant enhancement of the knowledge in understanding the phenomenology, identifying the psychopathology, diagnosing common mental disorder and selecting the psychotropic medication in the bivariate analysis. When the possible confounders of level of training in paediatrics and number of years spent as paediatrician were controlled, in addition to the above areas of adolescent psychiatry, the diagnostic ability involving multiple psychological concepts also gained significance. However, both in the bivariate and multivariate analyses, the ability to refer to appropriate psychotherapy remained unchanged after the workshop. Conclusions This workshop was effective in enhancing the adolescent psychiatry knowledge of paediatricians. Such workshops could strengthen paediatricians in addressing the priority mental health disorders at the primary-care level in countries with low-human resource for health as advocated by the World Health Organization. However, it remains to be seen if this acquisition of adolescent psychiatry knowledge results in enhancing their adolescent psychiatry practice. PMID:20167069

  20. Family Workshops

    ERIC Educational Resources Information Center

    Bennett, Dave; Rees-Jones, Tanny

    1978-01-01

    A Family Workshop is an informal, multidisciplined educational program for adults and children, organized by a team of teachers. This article discusses the Lavender Hill Family Workshop, one of many, which attempts to provide education in various subject areas for adults and for children while also integrating both objectives in order to educate…

  1. JC polyomavirus mutants escape antibody-mediated neutralization.

    PubMed

    Ray, Upasana; Cinque, Paola; Gerevini, Simonetta; Longo, Valeria; Lazzarin, Adriano; Schippling, Sven; Martin, Roland; Buck, Christopher B; Pastrana, Diana V

    2015-09-23

    JC polyomavirus (JCV) persistently infects the urinary tract of most adults. Under conditions of immune impairment, JCV causes an opportunistic brain disease, progressive multifocal leukoencephalopathy (PML). JCV strains found in the cerebrospinal fluid of PML patients contain distinctive mutations in surface loops of the major capsid protein, VP1. We hypothesized that VP1 mutations might allow the virus to evade antibody-mediated neutralization. Consistent with this hypothesis, neutralization serology revealed that plasma samples from PML patients neutralized wild-type JCV strains but failed to neutralize patient-cognate PML-mutant JCV strains. This contrasted with serological results for healthy individuals, most of whom robustly cross-neutralized all tested JCV variants. Mice administered a JCV virus-like particle (VLP) vaccine initially showed neutralizing "blind spots" (akin to those observed in PML patients) that closed after booster immunization. A PML patient administered an experimental JCV VLP vaccine likewise showed markedly increased neutralizing titer against her cognate PML-mutant JCV. The results indicate that deficient humoral immunity is a common aspect of PML pathogenesis and that vaccination may overcome this humoral deficiency. Thus, vaccination with JCV VLPs might prevent the development of PML.

  2. Hydroxyproline in the major capsid protein VP1 of polyomavirus

    SciTech Connect

    Ludlow, J.W.; Consigli, R.A.

    1989-06-01

    Amino acid analysis of (/sup 3/H)proline-labeled polyomavirus major capsid protein VP1 by two-dimensional paper chromatography of the acid-hydrolyzed protein revealed the presence of /sup 3/H-labeled hydroxyproline. Addition of the proline analog L-azetidine-2-carboxylic acid to infected mouse kidney cell cultures prevented or greatly reduced hydroxylation of proline in VP1. Immunofluorescence analysis performed on infected cells over a time course of analog addition revealed that virus proteins were synthesized but that transport from the cytoplasm to the nucleus was impeded. A reduction in the assembly of progeny virions demonstrated by CsCl gradient purification of virus from (/sup 35/S)methionine-labeled infected cell cultures was found to correlate with the time of analog addition. These results suggest that incorporation of this proline analog into VP1, accompanied by reduction of the hydroxyproline content of the protein, influences the amount of virus progeny produced by affecting transport of VP1 to the cell nucleus for assembly into virus particles.

  3. Continuing to Build a Community Consensus on the Future of Human Space Flight: Report of the Fourth Community Workshop on Achievability and Sustainability of Human Exploration of Mars (AM IV)

    NASA Technical Reports Server (NTRS)

    Thronson, Harley A.; Baker, John; Beaty, David; Carberry, Chris; Craig, Mark; Davis, Richard M.; Drake, Bret G.; Cassady, Joseph; Hays, Lindsay; Hoffman, Stephen J.; Mason, Lee S.

    2016-01-01

    To continue to build broadly based consensus on the future of human space exploration, the Fourth Community Workshop on Achievability and Sustainability of Human Exploration of Mars (AM IV), organized by Explore Mars, Inc. and the American Astronautical Society, was held at the Double Tree Inn in Monrovia, CA., December 68, 2016. Approximately 60 invited professionals from the industrial and commercial sectors, academia, and NASA, along with international colleagues, participated in the workshop. These individuals were chosen to be representative of the breadth of interests in astronaut and robotic Mars exploration.

  4. Polyomavirus Infection in Gouldian Finches (Erythrura gouldiae) and Other Pet Birds of the Family Estrildidae.

    PubMed

    Circella, E; Caroli, A; Marino, M; Legretto, M; Pugliese, N; Bozzo, G; Cocciolo, G; Dibari, D; Camarda, A

    2017-02-18

    A syndrome characterized by apathy, diarrhoea and high mortality of nestlings was observed in a flock of pet birds of the family Estrildidae. Enlargement of the liver, pulmonary congestion and urate accretions in the kidney were observed. Microscopically, there was glomerular atrophy, oedema and congestion of the lungs and necrosis and fibrosis of the liver. Cowdry type B intranuclear inclusion bodies were detected in the tissues. Polyomavirus was detected by polymerase chain reaction. The entire genome of the virus was amplified and sequenced, revealing 99 % identity to the sequence of finch polyomavirus isolated from the Eurasian bullfinch (family Fringillidae).

  5. Chromosome 19 International Workshop

    SciTech Connect

    Pericak-Vance, M.A. . Medical Center); Ropers, H.H. . Dept. of Human Genetics); Carrano, A.J. )

    1993-01-04

    The Second International Workshop on Human Chromosome 19 was hosted on January 25 and 26, 1992, by the Department of Human Genetics, University Hospital Nijmegen, The Netherlands, at the 'Meerdal Conference Center'. The workshop was supported by a grant from the European Community obtained through HUGO, the Dutch Research Organization (NWO) and the Muscular Dystrophy Association (MDA). Travel support for American participants was provided by the Department of Energy. The goals of this workshop were to produce genetic, physical and integrated maps of chromosome 19, to identify inconsistencies and gaps, and to discuss and exchange resources and techniques available for the completion of these maps. The second day of the meeting was largely devoted to region or disease specific efforts. In particular, the meeting served as a platform for assessing and discussing the recent progress made into the molecular elucidation of myotonic dystrophy.

  6. NEH Western Humanities Conference Workshop (Asilomar, California, August 2-5, 1977).

    ERIC Educational Resources Information Center

    1977

    Descriptions of nine humanities programs supported by the National Endowment for the Humanities at two- and four-year colleges are presented in this document. These include a 15-hour team taught interdisciplinary program for technology students at Abraham Baldwin Agricultural College (Georgia); the Bay Area Writing Project, aimed at strengthening…

  7. Conserved archetypal configuration of the transcriptional control region during the course of BK polyomavirus evolution.

    PubMed

    Yogo, Yoshiaki; Zhong, Shan; Xu, Yawei; Zhu, Mengyun; Chao, Yuegen; Sugimoto, Chie; Ikegaya, Hiroshi; Shibuya, Ayako; Kitamura, Tadaichi

    2008-08-01

    BK polyomavirus (BKV) is widespread among humans, asymptomatically infecting children and then persisting in renal tissue. The transcriptional control region (TCR) of the BKV genome is variable among clinical isolates. Thus, archetypal TCRs with a common basic configuration generally occur in BKV isolates from the urine of immunocompromised patients, but rearranged TCRs that possibly arise from the archetypal configuration have also been detected in clinical specimens. To examine the hypothesis that archetypal strains represent wild-type strains circulating in the human population (the archetype hypothesis), we analysed 145 complete viral genomes amplified directly from the urine of non-immunocompromised individuals worldwide. These genomes included 82, three, two and 58 sequences classified as belonging to subtypes I, II, III and IV, respectively. Rearranged TCRs with long duplications or deletions were detected from two subtype I and two subtype IV genomes, but not from the other 141 genomes (thus, the TCRs of these genomes were judged to be archetypal). The variations in the archetypal TCRs were nucleotide substitutions and single-nucleotide deletions, most of which were unique to particular subtypes or subgroups. We confirmed that the four complete BKV genomes with rearranged TCRs did not form a unique lineage on a phylogenetic tree. Collectively, the findings demonstrate that the archetypal TCR configuration has been conserved during the evolution of BKV, providing support for the archetype hypothesis. Additionally, we suggest that 'archetype' should be used as a conceptual term that denotes a prototypical structure that can generate various rearranged TCRs during viral growth in vivo and in vitro.

  8. Potential human health effects of acid rain: report of a workshop

    PubMed Central

    Goyer, Robert A.; Bachmann, John; Clarkson, Thomas W.; Ferris, Benjamin G.; Graham, Judith; Mushak, Paul; Perl, Daniel P.; Rall, David P.; Schlesinger, Richard; Sharpe, William; Wood, John M.

    1985-01-01

    This report summarizes the potential impact of the acid precipitation phenomenon on human health. There are two major components to this phenomenon: the predepositional phase, during which there is direct human exposure to acidic substances from ambient air, and the post-depositional phase, in which the deposition of acid materials on water and soil results in the mobilization, transport, and even chemical transformation of toxic metals. Acidification increases bioconversion of mercury to methylmercury, which accumulates in fish, increasing the risk to toxicity in people who eat fish. Increase in water and soil content of lead and cadmium increases human exposure to these metals which become additive to other sources presently under regulatory control. The potential adverse health effects of increased human exposure to aluminum is not known at the present time. PMID:3896772

  9. Risk of squamous cell skin cancer after organ transplant associated with antibodies to cutaneous papillomaviruses, polyomaviruses, and TMC6/8 (EVER1/2) variants.

    PubMed

    Madeleine, Margaret M; Carter, Joseph J; Johnson, Lisa G; Wipf, Gregory C; Davis, Connie; Berg, Daniel; Nelson, Karen; Daling, Janet R; Schwartz, Stephen M; Galloway, Denise A

    2014-10-01

    Squamous cell skin cancer (SCSC) disproportionately affects organ transplant recipients, and may be related to increased viral replication in the setting of immune suppression. We conducted a nested case-control study among transplant recipients to determine whether SCSC is associated with antibodies to cutaneous human papillomaviruses (HPV), to genes associated with a rare genetic susceptibility to HPV (TMC6/TMC8), or to human polyomaviruses (HPyV). Cases (n = 149) had histologically confirmed SCSC, and controls (n = 290) were individually matched to cases on time since transplant, type of transplant, gender, and race. All subjects had serum drawn immediately prior to transplant surgery. Antibodies to 25 cutaneous HPVs and six HPyVs were assayed by detection of binding to virus-like particles, and 11 TMC6/8 variants were genotyped. After correction for multiple comparisons, only antibodies to HPV37 were associated with SCSC (OR 2.0, 95% CI 1.2-3.4). Common genetic variants of TMC6/8 were not associated with SCSC, but three variants in TMC8 (rs12452890, rs412611, and rs7208422) were associated with greater seropositivity for species 2 betapapillomaviruses among controls. This study suggests that some betaHPVs, but not polyomaviruses, may play a role in the excess risk of SCSC among transplant recipients.

  10. Planetary Protection Knowledge Gaps for Human Extraterrestrial Missions Workshop Booklet - 2015

    NASA Technical Reports Server (NTRS)

    Fonda, Mark L.

    2015-01-01

    Although NASA's preparations for the Apollo lunar missions had only a limited time to consider issues associated with the protection of the Moon from biological contamination and the quarantine of the astronauts returning to Earth, they learned many valuable lessons (both positive and negative) in the process. As such, those efforts represent the baseline of planetary protection preparations for sending humans to Mars. Neither the post-Apollo experience or the Shuttle and other follow-on missions of either the US or Russian human spaceflight programs could add many additional insights to that baseline. Current mission designers have had the intervening four decades for their consideration, and in that time there has been much learned about human-associated microbes, about Mars, and about humans in space that has helped prepare us for a broad spectrum of considerations regarding potential biological contamination in human Mars missions and how to control it. This paper will review the approaches used in getting this far, and highlight some implications of this history for the future development of planetary protection provisions for human missions to Mars. The role of NASA and ESA's planetary protection offices, and the aegis of COSPAR have been particularly important in the ongoing process.

  11. Course/Workshop Complementarity

    ERIC Educational Resources Information Center

    Kane, Dan

    1976-01-01

    This paper discusses the law-related studies provided in a human ecology degree program. The studies involve workshops which are project-oriented experiences and courses which provide skills and knowledge. The program emphasizes law relating to land use management, small business enterprises, consumer protection, real estate, and family. (MR)

  12. Development of a DNA vaccine targeting Merkel cell polyomavirus.

    PubMed

    Zeng, Qi; Gomez, Bianca P; Viscidi, Raphael P; Peng, Shiwen; He, Liangmei; Ma, Barbara; Wu, T-C; Hung, Chien-Fu

    2012-02-08

    Merkel cell carcinoma (MCC) is a rare but devastating skin disease that is increasing in incidence within the United States. The poor prognosis of MCC patients and limited understanding of MCC pathogenesis warrants innovative treatments to control MCC. Several lines of evidence have pointed to Merkel cell polyomavirus (MCPyV) as the etiological agent of MCC. In particular, the amino terminus of MCPyV large T antigen (LT) (aa1-258) is expressed in all MCPyV-positive tumors and plays an important role in MCC oncogenesis, rendering it an ideal therapeutic target for vaccination. In the current study, we developed a DNA vaccine encoding MCPyV LT aa1-258 (pcDNA3-LT). Within our pcDNA3-LT DNA vaccine, we identified that MCPyV LT aa136-160 likely contains an LT-specific CD4+ T helper epitope. We have also created an LT-expressing B16/LT tumor model using B16, a murine melanoma cell line, to characterize the potency of our DNA vaccine. Using this tumorigenic B16/LT tumor model, we found that pcDNA3-LT DNA vaccine generates antitumor effects mainly mediated by CD4+ T cells against B16/LT tumors in vaccinated C57BL/6 mice. Thus, immunotherapy using pcDNA3-LT DNA vaccine may represent a promising approach for the control of MCPyV-associated lesions. The B16/LT tumor model further serves as a useful model for testing various vaccine strategies against MCC.

  13. Workshop Reports

    NASA Astrophysics Data System (ADS)

    2012-04-01

    19 Workshops were held during IAU S285. 15 submitted reports of the discussions that took place, while for the remaining 4 we have reproduced the summaries that were available on our wiki prior to the Symposium.

  14. Rapid Detection of Trichodysplasia Spinulosa-Associated Polyomavirus in Skin Biopsy Specimen

    PubMed Central

    Urbano, Paulo Roberto P.; Pannuti, Cláudio Sérgio; Pierrotti, Ligia C.; David-Neto, Elias

    2014-01-01

    Trichodysplasia spinulosa-associated polyomavirus (TSV) is responsible for a rare skin cancer. Using metagenomic approaches, we determined the complete genome sequence of a TSV first detected in Brazil in spicules of an immunocompromised patient suspected to have trichodysplasia spinulosa. PMID:25059864

  15. Avian Polyomavirus Genome Sequences Recovered from Parrots in Captive Breeding Facilities in Poland

    PubMed Central

    Dayaram, Anisha; Piasecki, Tomasz; Chrząstek, Klaudia; White, Robyn; Julian, Laurel; van Bysterveldt, Katherine

    2015-01-01

    Eight genomes of avian polyomaviruses (APVs) were recovered and sequenced from deceased Psittacula eupatria, Psittacula krameri, and Melopsittacus undulatus from various breeding facilities in Poland. Of these APV-positive samples, six had previously tested positive for beak and feather disease virus (BFDV) and/or parrot hepatitis B virus (PHBV). PMID:26404592

  16. Merkel Cell Polyomavirus DNA in Respiratory Specimens from Children and Adults

    PubMed Central

    Lambert, Stephen B.; Whiley, David M.; Nissen, Michael D.; Sloots, Theo P.

    2009-01-01

    Merkel cell polyomavirus (MCPyV) DNA was detected in 7 (1.3%) of 526 respiratory tract samples from patients in Australia with upper or lower respiratory tract symptoms. Partial T antigen and major capsid protein sequences of MCPyV identified in respiratory secretions showed high homology (99%–100%) to those found in Merkel cell carcinoma. PMID:19239774

  17. Nucleotides in the polyomavirus enhancer that control viral transcription and DNA replication.

    PubMed Central

    Tang, W J; Berger, S L; Triezenberg, S J; Folk, W R

    1987-01-01

    The polyomavirus enhancer is required in cis for high-level expression of the viral early region and for replication of the viral genome. We introduced multiple mutations in the enhancer which reduced transcription and DNA replication. Polyomaviruses with these mutant enhancers formed very small plaques in whole mouse embryo cells. Revertants of the viral mutants were isolated and characterized. Reversion occurred by any of the following events: restoration of guanosines at nucleotide (nt) 5134 and nt 5140 within the adenovirus 5 E1A enhancer core AGGAAGTGACT; acquisition of an A----G mutation at nt 5258, which is the same mutation that enables polyomavirus to grow in embryonal carcinoma F9 cells; duplication of mutated sequences between nt 5146 and 5292 (including sequences homologous with immunoglobulin G, simian virus 40, and bovine papillomavirus enhancer elements). Reversion restored both the replicative and transcriptional functions of the viruses. Revertants that acquired the F9 mutation at nt 5258 grew at least 20-fold better than the original mutant in whole mouse embryo cells, but replicated only marginally better than the original mutant in 3T6 cells. Viruses with a reversion of the mutation at nt 5140 replicated equally well in both types of cells. Since individual nucleotides in the polyomavirus enhancer simultaneously altered DNA replication and transcription in specific cell types, it is likely that these processes rely upon a common element, such as an enhancer-binding protein. Images PMID:3037332

  18. Polyomavirus (BK)-associated pleomorphic giant cell carcinoma of the urinary bladder: a case report.

    PubMed

    Alexiev, Borislav A; Papadimitriou, John C; Chai, Toby C; Ramos, Emilio; Staats, Paul N; Drachenberg, Cinthia B

    2013-04-01

    This report describes the morphological features of a pleomorphic giant cell carcinoma with focal trophoblastic differentiation of the urinary bladder in a male, 12 years post living related donor renal transplant. The voided urine cytology demonstrated rare decoy cells admixed with markedly atypical urothelial cell clusters, papillae and giant cells. Cystoprostatectomy demonstrated a nodular mass involving the trigone and right lateral-posterior wall, adjacent to the ureteral orifice. Hematoxylin-eosin stained sections showed two synchronous malignancies: (a) pleomorphic giant cell carcinoma with focal trophoblastic differentiation of the urinary bladder, metastatic to the omentum and (b) prostatic adenocarcinoma, Gleason score 3+4=7, involving the right prostate lobe. Strong diffuse expression of polyomavirus large T antigen was demonstrated in the primary and metastatic pleomorphic giant cell carcinoma, supporting a possible role for polyomavirus (BK) in the oncogenetic pathway. The prostatic adenocarcinoma was negative for polyomavirus large T antigen. Our findings of p63, CK7 and CK903 expression in pleomorphic giant cell carcinoma suggest that the tumor is of urothelial derivation. This is the first report describing the morphological features of urinary bladder pleomorphic giant cell carcinoma with trophoblastic differentiation, positive for polyomavirus large T antigen, arising in the background of BKV reactivation.

  19. Polyomavirus detection in multiple sclerosis patients under natalizumab therapy: Profile and frequency of urinary shedding.

    PubMed

    Nali, Luiz Henrique; Fink, Maria Cristina; do Olival, Guilherme S; Moraes, Lenira; Callegaro, Dagoberto; Tilbery, Charles Peter; Vidal, Jose Ernesto; Sumita, Laura Masami; de Oliveira, Augusto C Penalva; Romano, Camila M

    2017-03-01

    Patients undergoing Natalizumab (NTZ) therapy are at risk of progressive multifocal leukoencephalopathy (PML). Besides John Cunningham virus (JCV), BK polyomavirus might represent an additional concern for such patients since it can also infect CNS cells. Currently, data regarding the presence of anti-JCV antibodies added to previous immunosuppressive therapy and prolonged NTZ therapy has been used to classify patients at risk of developing PML. Here, we investigated the profile shedding of JCV and BKV in multiple sclerosis (MS) patients during treatment with NTZ. Serial blood and urine samples from 97 MS patients receiving either NTZ or β-interferon were investigated for polyomavirus shedding. While all blood samples tested negative, 36% of the patients shed polyomavirus in the urine in at least one time point. From these, 21.7%, 9.3%, and 5.1% shed JCV, BKV, and both polyomavirus, respectively. No difference was observed between the rates of urinary shedding of patients treated with NTZ (38.9%) and patients treated with other drugs (34.5%), also no PML event was diagnosed during the follow-up. Therefore, urinary shedding might not be interfered by therapy condition. In our study, we also observed 14/27 (52%) of anti-JCV antibodies prevalence, and nearly half of them (42%) did not present any event of urinary shedding during the follow-up. J. Med. Virol. 89:528-534, 2017. © 2016 Wiley Periodicals, Inc.

  20. Trichodysplasia spinulosa: a benign adnexal proliferation with follicular differentiation associated with polyomavirus.

    PubMed

    Lee, Yvonne Y; Tucker, Simon C; Prow, Natalie A; Setoh, Yin Xiang; Banney, Leith A

    2014-05-01

    Trichodysplasia spinulosa is a rare polyomavirus-associated cutaneous eruption occurring in the setting of immunosuppression. Clinically it is characterised by multiple centrofacial folliculocentric papules with spinous protuberances. The histopathology is distinct and treatment with antiviral agents appears to be the most effective.

  1. The Cre/loxP recombination system for production of infectious mouse polyomavirus.

    PubMed

    Hron, Tomáš; Spanielová, Hana; Suchanová, Jiřina; Forstová, Jitka

    2013-09-01

    Murine polyomavirus mutants are frequently produced for experimental as well as therapy purposes. Commonly used methods for preparation of mutant viral genomes from recombinant vectors are laborious and give variable yields and quality. We describe an efficient and reproducible Cre/loxP-mediated recombination system that generates polyomavirus genomes from recombinant plasmid in vivo. We designed and constructed two variants of recombinant vectors containing the wild-type polyomavirus genome flanked by loxP homologous sites. The loxP sites were introduced either into the intronic region of early genes or between the two poly(A) signal sites of convergent transcriptional units. After cotransfection of the recombinant plasmids with the Cre-expressing vector into mouse 3T6 cells, we obtained infectious virus from the genome variant containing loxP site in the intronic region, but we failed to isolate any infectious virus from the viral genome containing loxP site between poly(A) signals. We show that the Cre/loxP-based method of polyomavirus production is simple, expedient, and reproducible and works with satisfactory efficiency.

  2. MAKING SENSE OF HUMAN BIOMONITORING DATA: FINDINGS AND RECOMMENDATIONS OF A WORKSHOP

    EPA Science Inventory

    The ability to measure chemicals in humans (often termed biomonitoring) is far outpacing the ability to reliably interpret these data for public health purposes, creating a major knowledge gap. Until this gap is filled, the great promise of routinely using biomonitoring data to s...

  3. Human-Centred Design Workshops in Collaborative Strategic Design Projects: An Educational and Professional Comparison

    ERIC Educational Resources Information Center

    Liem, Andre; Sanders, Elizabeth B.-N.

    2013-01-01

    It has been found that the implementation of Human-centred Design (HCD) methods in the Fuzzy Front-End is not likely to lead to diversification in educational product planning exercises, where time lines are short and executors lack experience. Companies, interested to collaborate with Master-level Industrial Design students on strategic design…

  4. Antiviral effects of artesunate on JC polyomavirus replication in COS-7 cells.

    PubMed

    Sharma, Biswa Nath; Marschall, Manfred; Rinaldo, Christine Hanssen

    2014-11-01

    The human JC polyomavirus (JCPyV) causes the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML). A growing number of patients with induced or acquired immunosuppression are at risk for infection, and no effective antiviral therapy is presently available. The widely used antimalarial drug artesunate has shown broad antiviral activity in vitro but limited clinical success. The aim of this study was to investigate the effect of artesunate on JCPyV replication in vitro. The permissivity for JCPyV MAD-4 was first compared in four cell lines, and the monkey kidney cell line COS-7 was selected. Artesunate caused a concentration-dependent decrease in the extracellular JCPyV DNA load 96 h postinfection, with a 50% effective concentration (EC50) of 2.9 μM. This effect correlated with a decreased expression of capsid protein VP1 and a reduced release of infectious viral progeny. For concentrations of <20 μM, transient reductions in cellular DNA replication and proliferation were seen, while for higher concentrations, some cytotoxicity was detected. A selective index of 16.6 was found when cytotoxicity was calculated based on cellular DNA replication in the mock-infected cells, but interestingly, cellular DNA replication in the JCPyV-infected cells was more strongly affected. In conclusion, artesunate is efficacious in inhibiting JCPyV replication at micromolar concentrations, which are achievable in plasma. The inhibition at EC50 probably reflects an effect on cellular proteins and involves transient cytostatic effects. Our results, together with the favorable distribution of the active metabolite dihydroartemisinin to the central nervous system, suggest a potential use for artesunate in patients with PML.

  5. JC polyomavirus attachment, entry, and trafficking: unlocking the keys to a fatal infection.

    PubMed

    Maginnis, Melissa S; Nelson, Christian D S; Atwood, Walter J

    2015-12-01

    The human JC polyomavirus (JCPyV) causes a lifelong persistent infection in the reno-urinary tract in the majority of the adult population worldwide. In healthy individuals, infection is asymptomatic, while in immunocompromised individuals, the virus can spread to the central nervous system and cause a fatal demyelinating disease known as progressive multifocal leukoencephalopathy (PML). There are currently very few treatment options for this rapidly progressing and devastating disease. Understanding the basic biology of JCPyV-host cell interactions is critical for the development of therapeutic strategies to prevent or treat PML. Research in our laboratory has focused on gaining a detailed mechanistic understanding of the initial steps in the JCPyV life cycle in order to define how JCPyV selectively targets cells in the kidney and brain. JCPyV requires sialic acids to attach to host cells and initiate infection, and JCPyV demonstrates specificity for the oligosaccharide lactoseries tetrasaccharide c (LSTc) with an α2,6-linked sialic acid. Following viral attachment, JCPyV entry is facilitated by the 5-hydroxytryptamine (5-HT)2 family of serotonin receptors via clathrin-dependent endocytosis. JCPyV then undergoes retrograde transport to the endoplasmic reticulum (ER) where viral disassembly begins. A novel retrograde transport inhibitor termed Retro-2(cycl) prevents trafficking of JCPyV to the ER and inhibits both initial virus infection and infectious spread in cell culture. Understanding the molecular mechanisms by which JCPyV establishes infection will open up new avenues for the prevention or treatment of virus-induced disease.

  6. JC Polyomavirus Attachment, Entry, and Trafficking: Unlocking the Keys to a Fatal Infection

    PubMed Central

    Maginnis, Melissa S.; Nelson, Christian D.S.; Atwood, Walter J.

    2014-01-01

    The human JC polyomavirus (JCPyV) causes a lifelong persistent infection in the reno-urinary tract in the majority of the adult population worldwide. In healthy individuals infection is asymptomatic, while in immunocompromised individuals the virus can spread to the central nervous system and cause a fatal demyelinating disease known as progressive multifocal leukoencephalopathy (PML). There are currently very few treatment options for this rapidly progressing and devastating disease. Understanding the basic biology of JCPyV-host cell interactions is critical for the development of therapeutic strategies to prevent or treat PML. Research in our laboratory has focused on gaining a detailed mechanistic understanding of the initial steps in the JCPyV life cycle in order to define how JCPyV selectively targets cells in the kidney and brain. JCPyV requires sialic acids to attach to host cells and initiate infection, and JCPyV demonstrates specificity for the oligosaccharide lactoseries tetrasaccharide c (LSTc) with an α2,6-linked sialic acid. Following viral attachment, JCPyV entry is facilitated by the 5-hydroxytryptamine (5-HT)2 family of serotonin receptors via clathrin-dependent endocytosis. JCPyV then undergoes retrograde transport to the endoplasmic reticulum (ER) where viral disassembly begins. A novel retrograde transport inhibitor termed Retro-2cycl prevents trafficking of JCPyV to the ER and inhibits both initial virus infection and infectious spread in cell culture. Understanding the molecular mechanisms by which JCPyV establishes infection will open up new avenues for the prevention or treatment of virus-induced disease. PMID:25078361

  7. Workshop on Numerical Analysis of Human and Surrogate Response to Accelerative Loading

    DTIC Science & Technology

    2014-05-01

    human systems. For blast and ballistic applications, both Lagrangian and Eulerian techniques are essential to capture various fluid-structural... ballistic rates. The primary reason for the lack of data is that most of the high-rate experimental techniques were developed for stiffer and stronger...accelerative or terminal ballistic loading conditions. This is accomplished by adding numerical methods for Lagrangian field behavior, separate

  8. DOT/FAA Human Factors Workshop on Aviation. Transcript. Volume II.

    DTIC Science & Technology

    1980-11-25

    into our judgements . We participate in other areas, in evacuation systems, interior design, things like that, whole broad range of activities including...human factors people in that generation or that era spent too much time worrying about displays and controls in his judgement . He said generally we...pilots, perhaps more often than some of them, generally line pilots to come in and fly and give their judgements on the concepts that we have displayed

  9. 75 FR 58411 - Center for Veterinary Medicine eSubmitter Workshop; Public Workshop; Request for Comments

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-24

    ... HUMAN SERVICES Food and Drug Administration Center for Veterinary Medicine eSubmitter Workshop; Public...: ``Center for Veterinary Medicine (CVM) eSubmitter Workshop.'' The purpose of the public workshop is to..., Center for Veterinary Medicine (HFV-100), Food and Drug Administration, 7520 Standish Pl., Rockville,...

  10. The structure of avian polyomavirus reveals variably sized capsids, non-conserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4

    SciTech Connect

    Shen, Peter S.; Enderlein, Dirk; Nelson, Christian D.S.; Carter, Weston S.; Kawano, Masaaki; Xing Li; Swenson, Robert D.; Olson, Norman H.; Baker, Timothy S.; Cheng, R. Holland; Atwood, Walter J.; Johne, Reimar; Belnap, David M.

    2011-03-01

    Avian polyomavirus (APV) causes a fatal, multi-organ disease among several bird species. Using cryogenic electron microscopy and other biochemical techniques, we investigated the structure of APV and compared it to that of mammalian polyomaviruses, particularly JC polyomavirus and simian virus 40. The structure of the pentameric major capsid protein (VP1) is mostly conserved; however, APV VP1 has a unique, truncated C-terminus that eliminates an intercapsomere-connecting {beta}-hairpin observed in other polyomaviruses. We postulate that the terminal {beta}-hairpin locks other polyomavirus capsids in a stable conformation and that absence of the hairpin leads to the observed capsid size variation in APV. Plug-like density features were observed at the base of the VP1 pentamers, consistent with the known location of minor capsid proteins VP2 and VP3. However, the plug density is more prominent in APV and may include VP4, a minor capsid protein unique to bird polyomaviruses.

  11. Characterization of a novel polyomavirus isolated from a fibroma on the trunk of an African elephant (Loxodonta africana).

    PubMed

    Stevens, Hans; Bertelsen, Mads Frost; Sijmons, Steven; Van Ranst, Marc; Maes, Piet

    2013-01-01

    Viruses of the family Polyomaviridae infect a wide variety of avian and mammalian hosts with a broad spectrum of outcomes including asymptomatic infection, acute systemic disease, and tumor induction. In this study a novel polyomavirus, the African elephant polyomavirus 1 (AelPyV-1) found in a protruding hyperplastic fibrous lesion on the trunk of an African elephant (Loxodonta africana) was characterized. The AelPyV-1 genome is 5722 bp in size and is one of the largest polyomaviruses characterized to date. Analysis of the AelPyV-1 genome reveals five putative open-reading frames coding for the classic small and large T antigens in the early region, and the VP1, VP2 and VP3 capsid proteins in the late region. In the area preceding the VP2 start codon three putative open-reading frames, possibly coding for an agnoprotein, could be localized. A regulatory, non-coding region separates the 2 coding regions. Unique for polyomaviruses is the presence of a second 854 bp long non-coding region between the end of the early region and the end of the late region. Based on maximum likelihood phylogenetic analyses of the large T antigen of the AelPyV-1 and 61 other polyomavirus sequences, AelPyV-1 clusters within a heterogeneous group of polyomaviruses that have been isolated from bats, new world primates and rodents.

  12. The functions of human saliva: A review sponsored by the World Workshop on Oral Medicine VI.

    PubMed

    Dawes, C; Pedersen, A M L; Villa, A; Ekström, J; Proctor, G B; Vissink, A; Aframian, D; McGowan, R; Aliko, A; Narayana, N; Sia, Y W; Joshi, R K; Jensen, S B; Kerr, A R; Wolff, A

    2015-06-01

    This narrative review of the functions of saliva was conducted in the PubMed, Embase and Web of Science databases. Additional references relevant to the topic were used, as our key words did not generate references which covered all known functions of saliva. These functions include maintaining a moist oral mucosa which is less susceptible to abrasion, and removal of micro-organisms, desquamated epithelial cells, leucocytes and food debris by swallowing. The mucins form a slimy coating on all surfaces in the mouth and act as a lubricant during such processes as mastication, formation of a food bolus, swallowing and speaking. Saliva provides the fluid in which solid tastants may dissolve and distributes tastants around the mouth to the locations of the taste buds. The hypotonic unstimulated saliva facilitates taste recognition. Salivary amylase is involved in digestion of starches. Saliva acts as a buffer to protect oral, pharyngeal and oesophageal mucosae from orally ingested acid or acid regurgitated from the stomach. Saliva protects the teeth against acid by contributing to the acquired enamel pellicle, which forms a renewable lubricant between opposing tooth surfaces, by being supersaturated with respect to tooth mineral, by containing bicarbonate as a buffer and urea and by facilitating clearance of acidic materials from the mouth. Saliva contains many antibacterial, antiviral and antifungal agents which modulate the oral microbial flora in different ways. Saliva also facilitates the healing of oral wounds. Clearly, saliva has many functions which are needed for proper protection and functioning of the human body.

  13. Host DNA Damage Response Factors Localize to Merkel Cell Polyomavirus DNA Replication Sites To Support Efficient Viral DNA Replication

    PubMed Central

    Tsang, Sabrina H.; Wang, Xin; Li, Jing; Buck, Christopher B.

    2014-01-01

    ABSTRACT Accumulating evidence indicates a role for Merkel cell polyomavirus (MCPyV) in the development of Merkel cell carcinoma (MCC), making MCPyV the first polyomavirus to be clearly associated with human cancer. With the high prevalence of MCPyV infection and the increasing amount of MCC diagnosis, there is a need to better understand the virus and its oncogenic potential. In this study, we examined the relationship between the host DNA damage response (DDR) and MCPyV replication. We found that components of the ATM- and ATR-mediated DDR pathways accumulate in MCPyV large T antigen (LT)-positive nuclear foci in cells infected with native MCPyV virions. To further study MCPyV replication, we employed our previously established system, in which recombinant MCPyV episomal DNA is autonomously replicated in cultured cells. Similar to native MCPyV infection, where both MCPyV origin and LT are present, the host DDR machinery colocalized with LT in distinct nuclear foci. Immunofluorescence in situ hybridization and bromodeoxyuridine (BrdU) incorporation analysis showed that these DDR proteins and MCPyV LT in fact colocalized at the actively replicating MCPyV replication complexes, which were absent when a replication-defective LT mutant or an MCPyV-origin mutant was introduced in place of wild-type LT or wild-type viral origin. Inhibition of DDR kinases using chemical inhibitors and ATR/ATM small interfering RNA (siRNA) knockdown reduced MCPyV DNA replication without significantly affecting LT expression or the host cell cycle. This study demonstrates that these host DDR factors are important for MCPyV DNA replication, providing new insight into the host machinery involved in the MCPyV life cycle. IMPORTANCE MCPyV is the first polyomavirus to be clearly associated with human cancer. However, the MCPyV life cycle and its oncogenic mechanism remain poorly understood. In this report, we show that, in cells infected with native MCPyV virions, components of the ATM- and ATR

  14. Wordland Workshop.

    ERIC Educational Resources Information Center

    Perlish, Harvey Neil

    Can and should the preschool child learn to read? To answer this and related questions, a study was conducted to determine the effectiveness of a television program and parental home assistance in teaching reading skills to three-year-old children. For five days a week over a 39-week period, an experimental group watched "Wordland Workshop," a…

  15. Writers' Workshop.

    ERIC Educational Resources Information Center

    Sherris, Arieh

    1998-01-01

    Israeli 12th graders studying English as a Second Language benefit from writers' workshops where they compose written portfolios and learn to express themselves fluently in writing. Students write with paper and pen or work via the Internet. They write on selected issues and send letters and articles to various online and print journals and…

  16. Poetry Workshop.

    ERIC Educational Resources Information Center

    Janeczko, Paul B.

    2000-01-01

    This workshop offers activities to teach students about poetry. After describing haiku as a brief snapshot rather than a story, it explains how to teach poetry using an attached reproducible and poster. The tear-out reproducible sheet teaches students how to write their own haiku, offering a sample one as a model. The poster presents three sample…

  17. Teacher workshops

    NASA Technical Reports Server (NTRS)

    1993-01-01

    Education specialists with the NASA Educator Resource Center conduct a wide variety of workshops throughout the year to aid teachers and educators in coming up with new ideas to inspire their students and also in aiding in the integration of technology into their classrooms.

  18. Women's Workshop.

    ERIC Educational Resources Information Center

    Karelius, Karen

    The Women's Workshop Notebook is the tool used in the nine-week course designed for the mature woman returning to school at Antelope Valley College. The notebook exercises along with the group interaction and instruction stress the importance of personal assessment of strengths, weaknesses, dreams, deliberations and life history in…

  19. Winter Workshop.

    ERIC Educational Resources Information Center

    Council of Outdoor Educators of Quebec, Montreal.

    Materials on 11 topics presented at a winter workshop for Quebec outdoor educators have been compiled into this booklet. Action story, instant replay, shoe factory, sound and action, and find an object to fit the description are described and recommended as group dynamic activities. Directions for five games (Superlative Selection; Data…

  20. Low prevalence of Merkel cell polyomavirus with low viral loads in oral and maxillofacial tumours or tumour-like lesions from immunocompetent patients: Absence of Merkel cell polyomavirus-associated neoplasms

    PubMed Central

    TANIO, SHUNSUKE; MATSUSHITA, MICHIKO; KUWAMOTO, SATOSHI; HORIE, YASUSHI; KODANI, ISAMU; MURAKAMI, ICHIRO; RYOKE, KAZUO; HAYASHI, KAZUHIKO

    2015-01-01

    It was recently demonstrated that ~80% of Merkel cell carcinomas (MCCs) harbour a novel polyomavirus, Merkel cell polyomavirus (MCPyV). MCPyV has been detected in various human tissue samples. However, previous studies on the prevalence of MCPyV in oral tumours or tumour-like lesions are incomplete. To address this issue, we measured MCPyV DNA quantity using quantitative polymerase chain reaction (qPCR) in 327 oral tumours or tumour-like lesions and 54 jaw tumours or cyst lesions from 381 immunocompetent patients, as well as in 4 oral lesions from 4 immunosuppressed patients. qPCR revealed a low MCPyV prevalence (25/381, 6.6%) with low viral loads (0.00024-0.026 copies/cell) in oral and maxillofacial tumours and tumour-like lesions from immunocompetent patients. The prevalence was 7/176 (4.0%) in invasive squamous cell carcinomas (SCCs) [2/60 (3.33%) SCCs of the tongue, 4/52 (7.7%) SCCs of the gingiva and 1/19 (5.3%) SCCs of the floor of the mouth], 1/10 (10%) in dysplasias, 1/5 (20%) in adenocarcinomas, 2/13 (15.4%) in adenoid cystic carcinomas, 1/10 (10%) in non-Hodgkin's lymphomas, 3/10 (30%) in lipomas, 3/5 (60%) in neurofibromas, 1/3 (33.3%) in Schwannomas, 2/12 (16.7%) in Warthin's tumours, 2/11 (18.2%) in pyogenic granulomas, 1/14 (7.1%) in radicular cysts and 1/12 (8.3%) in ameloblastomas. The prevalence in lesions from immunosuppressed patients (1/4, 25%) was higher compared with that in lesions from immunocompetent patients (25/381, 6.6%), but the difference was not statistically significant. To the best of our knowledge, this study was the first to report prevalence data of MCPyV in tumours and cysts of the jaws (2/54, 3.7%). These data indicated absence of MCPyV-related tumours or tumour-like lesions in the oral cavity and jaws and suggested that the detected MCPyV DNA was derived from non-neoplastic background tissues with widespread low-level MCPyV infection. PMID:26807237

  1. Breaking Barriers: A Workshop Series in Human Relations Skills Based Upon Liberated Parents/Liberated Children. (Experimental Edition - Revised).

    ERIC Educational Resources Information Center

    Faber, Adele; Mazlish, Elaine

    This is one of a series of Education for Parenthood manuals developed for use in Salvation Army demonstration programs in parenthood education. This guide presents discussion plans for eight workshop sessions based on the book, "Liberated Parents/Liberated Children." The course was designed to help teenagers (1) become aware of typical negative…

  2. DOT/FAA Human Factors Workshop on Aviation (5th). Transcript Held at Oklahoma City, Oklahoma on 7-8 July 1981.

    DTIC Science & Technology

    1982-05-01

    1111" 1.0-2 Au11 I 12.0 NATIONAL BUREAU Of STANDARDS-1963-A -, b. e" • -O’-.."..’,o"- *-.. -,b -. ! . . ... .. ... °... tq . .,M ,..-o...TRANSPORTATION FEDERAL AVIATION ADMINISTRATION-’ Accession For NTTS GRA& I . . DTIC TAB Dv il~ ’ t .inn .. yCodes . and/or Dist !Special-’ Presented at the Mike...Fifth City, Oklahoma, on July 7-9, 1981. The Sixth Human Factors Workshop was held at the same facility on July 7 and 8, 1981./ I TABLE OF CONTENTS

  3. Safety and efficacy of an inactivated Carbopol-adjuvanted goose haemorrhagic polyomavirus vaccine for domestic geese.

    PubMed

    Gelfi, Jacqueline; Pappalardo, Michael; Claverys, Carine; Peralta, Brigitte; Guerin, Jean-Luc

    2010-04-01

    Haemorrhagic nephritis enteritis of the goose (HNEG) is an epizootic viral disease in domestic geese. The causal agent is a polyomavirus, namely goose haemorrhagic polyomavirus. To help control the disease, an inactivated vaccine was developed, based on viral particles produced in goose kidney cells. Viral material was quantified using real-time quantitative polymerase chain reaction, inactivated with beta-propiolactone and adjuvanted with Carbopol, an acrylic acid polymer. Carbopol proved to be more immunogenic than aluminium hydroxide and was totally safe when administered to young goslings and breeders alike. Carbopol-adjuvanted vaccine induced a high serological response. Moreover, goslings hatched from vaccinated breeders were protected against viral challenge, indicating that maternally-derived neutralizing antibodies (MDA) were efficiently transferred. MDA were still detectable 15 days post-hatch. Clinical trials will be necessary to accurately evaluate a vaccine-based HNEG control strategy under field conditions.

  4. Production and biomedical applications of virus-like particles derived from polyomaviruses.

    PubMed

    Teunissen, Erik A; de Raad, Markus; Mastrobattista, Enrico

    2013-11-28

    Virus-like particles (VLPs), aggregates of capsid proteins devoid of viral genetic material, show great promise in the fields of vaccine development and gene therapy. These particles spontaneously self-assemble after heterologous expression of viral structural proteins. This review will focus on the use of virus-like particles derived from polyomavirus capsid proteins. Since their first recombinant production 27 years ago these particles have been investigated for a myriad of biomedical applications. These virus-like particles are safe, easy to produce, can be loaded with a broad range of diverse cargoes and can be tailored for specific delivery or epitope presentation. We will highlight the structural characteristics of polyomavirus-derived VLPs and give an overview of their applications in diagnostics, vaccine development and gene delivery.

  5. Novel polyomavirus associated with brain tumors in free-ranging raccoons, western United States.

    PubMed

    Dela Cruz, Florante N; Giannitti, Federico; Li, Linlin; Woods, Leslie W; Del Valle, Luis; Delwart, Eric; Pesavento, Patricia A

    2013-01-01

    Tumors of any type are exceedingly rare in raccoons. High-grade brain tumors, consistently located in the frontal lobes and olfactory tracts, were detected in 10 raccoons during March 2010-May 2012 in California and Oregon, suggesting an emerging, infectious origin. We have identified a candidate etiologic agent, dubbed raccoon polyomavirus, that was present in the tumor tissue of all affected animals but not in tissues from 20 unaffected animals. Southern blot hybridization and rolling circle amplification showed the episomal viral genome in the tumors. The multifunctional nuclear protein large T-antigen was detectable by immunohistochemical analyses in a subset of neoplastic cells. Raccoon polyomavirus may contribute to the development of malignant brain tumors of raccoons.

  6. Merkel cell polyomavirus (MCPyV) in chronic lymphocytic leukemia/small lymphocytic lymphoma.

    PubMed

    Teman, Carolin J; Tripp, Sheryl R; Perkins, Sherrie L; Duncavage, Eric J

    2011-05-01

    Merkel cell polyomavirus (MCPyV) is a novel polyomavirus that shows a strong association with Merkel cell carcinoma (MCC). Recent studies have demonstrated MCPyV in some cases of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), a malignancy with a similar demographic as MCC. We tested for the presence of MCPyV by PCR and immunohistochemistry in 18 cases of CLL/SLL. Very low-level MCPyV DNA was detected in 33% of CLL/SLL cases by real-time PCR, but only one case demonstrated immunohistochemical positivity for MCPyV. MCPyV was not identified in 17 cases of follicular lymphoma, suggesting either that MCPyV is involved in CLL/SLL pathogenesis or that the immunodeficiency state of CLL/SLL induces low-level MCPyV reactivation.

  7. Structural optimization of a retrograde trafficking inhibitor that protects cells from infections by human polyoma- and papillomaviruses.

    PubMed

    Carney, Daniel W; Nelson, Christian D S; Ferris, Bennett D; Stevens, Julia P; Lipovsky, Alex; Kazakov, Teymur; DiMaio, Daniel; Atwood, Walter J; Sello, Jason K

    2014-09-01

    Human polyoma- and papillomaviruses are non-enveloped DNA viruses that cause severe pathologies and mortalities. Under circumstances of immunosuppression, JC polyomavirus causes a fatal demyelinating disease called progressive multifocal leukoencephalopathy (PML) and the BK polyomavirus is the etiological agent of polyomavirus-induced nephropathy and hemorrhagic cystitis. Human papillomavirus type 16, another non-enveloped DNA virus, is associated with the development of cancers in tissues like the uterine cervix and oropharynx. Currently, there are no approved drugs or vaccines to treat or prevent polyomavirus infections. We recently discovered that the small molecule Retro-2(cycl), an inhibitor of host retrograde trafficking, blocked infection by several human and monkey polyomaviruses. Here, we report diversity-oriented syntheses of Retro-2(cycl) and evaluation of the resulting analogs using an assay of human cell infections by JC polyomavirus. We defined structure-activity relationships and also discovered analogs with significantly improved potency as suppressors of human polyoma- and papillomavirus infection in vitro. Our findings represent an advance in the development of drug candidates that can broadly protect humans from non-enveloped DNA viruses and toxins that exploit retrograde trafficking as a means for cell entry.

  8. Structural Optimization of a Retrograde Trafficking Inhibitor that Protects Cells from Infections by Human Polyoma- and Papillomaviruses

    PubMed Central

    Carney, Daniel W.; Nelson, Christian D. S.; Ferris, Bennett D.; Stevens, Julia P.; Lipovsky, Alex; Kazakov, Temur; DiMaio, Daniel C.; Atwood, Walter J.; Sello, Jason K.

    2015-01-01

    Human polyoma- and papillomaviruses are non-enveloped DNA viruses that cause severe pathologies and mortalities. Under circumstances of immunosuppression, JC polyomavirus causes a fatal demyelinating disease called progressive multifocal leukoencephalopathy (PML) and the BK polyomavirus is the etiological agent of polyomavirus-induced nephropathy and hemorrhagic cystitis. Human papillomavirus type 16, another non-enveloped DNA virus, is associated with the development of cancers in tissues like the uterine cervix and oropharynx. Currently, there are no approved drugs or vaccines to treat or prevent polyomavirus infections. We recently discovered that the small molecule Retro-2cycl, an inhibitor of host retrograde trafficking, blocked infection by several human and monkey polyomaviruses. Here, we report diversity-oriented syntheses of Retro-2cycl and evaluation of the resulting analogs using an assay of human cell infections by JC polyomavirus. We defined structure-activity relationships and also discovered analogs with significantly improved potency as suppressors of human polyoma- and papillomavirus infection in vitro. Our findings represent an advance in the development of drug candidates that can broadly protect humans from non-enveloped DNA viruses and toxins that exploit retrograde trafficking as a means for cell entry. PMID:25087050

  9. AP1 enhances polyomavirus DNA replication by promoting T-antigen-mediated unwinding of DNA.

    PubMed Central

    Guo, W; Tang, W J; Bu, X; Bermudez, V; Martin, M; Folk, W R

    1996-01-01

    An early step in the initiation of polyomavirus DNA replication is viral large-T-antigen-mediated unwinding of the origin. We report that components of the AP1 transcription factor, Fos and Jun, interact with T antigen in vitro to enhance unwinding of the viral origin. This provides a biochemical basis for the capacity of AP1 to activate viral DNA replication in vivo. PMID:8763994

  10. Endemic Infection of Stranded Southern Sea Otters (Enhydra lutris nereis) with Novel Parvovirus, Polyomavirus, and Adenovirus.

    PubMed

    Siqueira, Juliana D; Ng, Terry; Miller, Melissa; Li, Linlin; Deng, Xutao; Dodd, Erin; Batac, Francesca; Delwart, Eric

    2017-02-13

    Over the past century, the southern sea otter (SSO; Enhydra lutris nereis) population has been slowly recovering from near extinction due to overharvest. The SSO is a threatened species under federal law and a fully protected species under California law. Through a multiagency collaborative program, stranded animals are rehabilitated and released, while deceased animals are necropsied and tissues are cryopreserved to facilitate scientific study. Here, we processed archival tissues to enrich particle-associated viral nucleic acids, which we randomly amplified and deeply sequenced to identify viral genomes through sequence similarities. Anelloviruses and endogenous retroviral sequences made up over 50% of observed viral sequences. Polyomavirus, parvovirus, and adenovirus sequences made up most of the remaining reads. We characterized and phylogenetically analyzed the full genome of sea otter (SO) polyomavirus 1 and the complete coding sequence of SO parvovirus 1 and found that the closest known viruses infect primates and domestic pigs ( Sus scrofa domesticus), respectively. We tested archived tissues from 69 stranded SO necropsied over 14 yr (2000-13) by PCR. Polyomavirus, parvovirus, and adenovirus infections were detected in 51, 61, and 29% of examined animals, respectively, with no significant increase in frequency over time, suggesting endemic infection. We found that 80% of tested SSO were infected with at least one of the three DNA viruses, whose tissue distribution we determined in 261 tissue samples. Parvovirus DNA was most frequently detected in mesenteric lymph node, polyomavirus DNA in spleen, and adenovirus DNA in multiple tissues (spleen, retropharyngeal and mesenteric lymph node, lung, and liver). This study describes the virome in tissues of a threatened species and shows that stranded SSO are frequently infected with multiple viruses, warranting future research to investigate associations between these infections and observed lesions.

  11. Identification of Merkel Cell Polyomavirus from a Patient with Acute Myeloid Leukemia

    PubMed Central

    Song, Y.

    2017-01-01

    ABSTRACT Merkel cell polyomavirus (MCPyV) is an oncogenic virus associated with Merkel cell carcinoma, an aggressive form of skin cancer with a high (>30%) mortality rate. The virus has a high incidence in patients with immunosuppressed conditions, such as AIDS or leukemia, or following organ transplantation. Here, we report the complete genomic sequence of MCPyV identified from a blood sample from a patient with acute myeloid leukemia. PMID:28104648

  12. Polyomavirus large T antigen binds symmetrical repeats at the viral origin in an asymmetrical manner.

    PubMed

    Harrison, Celia; Jiang, Tao; Banerjee, Pubali; Meinke, Gretchen; D'Abramo, Claudia M; Schaffhausen, Brian; Bohm, Andrew

    2013-12-01

    Polyomaviruses have repeating sequences at their origins of replication that bind the origin-binding domain of virus-encoded large T antigen. In murine polyomavirus, the central region of the origin contains four copies (P1 to P4) of the sequence G(A/G)GGC. They are arranged as a pair of inverted repeats with a 2-bp overlap between the repeats at the center. In contrast to simian virus 40 (SV40), where the repeats are nonoverlapping and all four repeats can be simultaneously occupied, the crystal structure of the four central murine polyomavirus sequence repeats in complex with the polyomavirus origin-binding domain reveals that only three of the four repeats (P1, P2, and P4) are occupied. Isothermal titration calorimetry confirms that the stoichiometry is the same in solution as in the crystal structure. Consistent with these results, mutation of the third repeat has little effect on DNA replication in vivo. Thus, the apparent 2-fold symmetry within the DNA repeats is not carried over to the protein-DNA complex. Flanking sequences, such as the AT-rich region, are known to be important for DNA replication. When the orientation of the central region was reversed with respect to these flanking regions, the origin was still able to replicate and the P3 sequence (now located at the P2 position with respect to the flanking regions) was again dispensable. This highlights the critical importance of the precise sequence of the region containing the pentamers in replication.

  13. Small and middle T antigens contribute to lytic and abortive polyomavirus infection

    SciTech Connect

    Tuerler, H.; Salomon, C.

    1985-02-01

    Using three different polyomavirus hr-t mutants and two polyomavirus mlT mutants, the authors studied induction of S-phase by mutants and wild-type virus in quiescent mouse kidney cells, mouse 3T6 cells, and FR 3T3 cells. At different times after infection, they measured the proportion of T-antigen-positive cells, the incorporation of (/sup 3/H)thymidine, the proportion of DNA-synthesizing cells, and the increase in total DNA, RNA, and protein content of the cultures. In permissive mouse cells, they also determined the amount of viral DNA and the proportion of viral capsid-producing cells. In polyomavirus hr-t mutant-infected cultures, the onset of host DNA replication was delayed by several hours, and a smaller proportion of T-antigen-positive cells entered S-phase than in wild-type-infected cultures. Of the two polyomavirus mlT mutants studied, dl-23 behaved similarly to wild-type virus in many, but not all, parameters tested. The poorly replicating but well-transforming mutant dl-8 was able to induce S-phase, and (in permissive cells) progeny virus production, in only about one-third of the T-antigen-positive cells. From the experiments, the authors concluded that mutations affecting small and middle T-antigen cause a reduction in the proportion of cells responding to virus infection and a prolongation of the early phase, i.e., the period before cells center S-phase. In hr-t mutant-infected mouse 3T6 cells, production of viral DNA was <10% of that in wild-type-infected cultures; low hr-t progeny production in 3T6 cells was therefore largely due to poor viral DNA replication.

  14. Polyomavirus Large T Antigen Binds Symmetrical Repeats at the Viral Origin in an Asymmetrical Manner

    PubMed Central

    Harrison, Celia; Jiang, Tao; Banerjee, Pubali; Meinke, Gretchen; D'Abramo, Claudia M.; Schaffhausen, Brian

    2013-01-01

    Polyomaviruses have repeating sequences at their origins of replication that bind the origin-binding domain of virus-encoded large T antigen. In murine polyomavirus, the central region of the origin contains four copies (P1 to P4) of the sequence G(A/G)GGC. They are arranged as a pair of inverted repeats with a 2-bp overlap between the repeats at the center. In contrast to simian virus 40 (SV40), where the repeats are nonoverlapping and all four repeats can be simultaneously occupied, the crystal structure of the four central murine polyomavirus sequence repeats in complex with the polyomavirus origin-binding domain reveals that only three of the four repeats (P1, P2, and P4) are occupied. Isothermal titration calorimetry confirms that the stoichiometry is the same in solution as in the crystal structure. Consistent with these results, mutation of the third repeat has little effect on DNA replication in vivo. Thus, the apparent 2-fold symmetry within the DNA repeats is not carried over to the protein-DNA complex. Flanking sequences, such as the AT-rich region, are known to be important for DNA replication. When the orientation of the central region was reversed with respect to these flanking regions, the origin was still able to replicate and the P3 sequence (now located at the P2 position with respect to the flanking regions) was again dispensable. This highlights the critical importance of the precise sequence of the region containing the pentamers in replication. PMID:24109229

  15. Activation of polyomavirus DNA replication by yeast GAL4 is dependent on its transcriptional activation domains.

    PubMed Central

    Bennett-Cook, E R; Hassell, J A

    1991-01-01

    The polyomavirus replication origin contains transcriptional regulatory sequences. To determine how these elements function in DNA replication, and to learn whether a common mechanism underlies the activation of transcription and DNA replication, we tested whether a well-characterized transcriptional activator, yeast GAL4, was capable of stimulating DNA replication and transcription in the same mammalian cell line. We observed that GAL4 activated polyomavirus DNA replication in mouse cells when its binding site was juxtaposed to the late border of the polyomavirus origin core. Synergistic activation of DNA replication was achieved by multimerization of the GAL4 binding site. Analysis of GAL4 mutant proteins, GAL4 hybrid proteins and mutants of the latter revealed that the activation domains of these transcriptional activators were required to stimulate DNA replication. In agreement with previously published data, the activation domains of GAL4 were also required to enhance transcription in the same mouse cell line. These observations implicate transcriptional activators in Py DNA replication and suggest that similar mechanisms govern the activation of transcription and DNA replication. Images PMID:1849079

  16. Cross-linking of a polyomavirus attachment protein to its mouse kidney cell receptor

    SciTech Connect

    Griffith, G.R.; Consigli, R.A.

    1986-06-01

    The authors used photoaffinity cross-linking with the heterobifunctional cross-linker N-hydroxysuccinimidyl 4-azidobenzoate (HSAB) to covalently link polyomavirus to a mouse kidney cell surface component. The virus-HSAB combination was adsorbed to the cells and then cross-linked and isolated in monopinocytotic vesicles from the cells after endocytosis. The cross-linked product was identified on sodium dodecyl sulfate-polyacrylamide gels by the presence of a new band carrying /sup 125/I-labeled virion protein with a higher molecular mass than the normal virion protein bands. A single new band, with an apparent molecular mass of 120 kilodaltons (120 kDa), was identified by this procedure. This band was formed only in the presence of the HSAB cross-linker when virions were bound to the cells. The band also copurified with cross-linked virions when virion-containing vesicles were treated with detergent to remove the cell membrane. Antibody treatments that blocked up to 100% of virus binding and internalization also blocked cross-linking, as measured by the formation of the 120-kDa band. The 120-kDa band was characterized by preparation of antibody against the excised band from the gel. These data have demonstrated that molecules of possible biological significance in the binding of polyomavirus to mouse kidney cells have been cross-linked and that cell surface molecules have been identified that may be characterized further for possible receptor function in polyomavirus attachment.

  17. Identification of an avian polyomavirus associated with Adélie penguins (Pygoscelis adeliae).

    PubMed

    Varsani, Arvind; Porzig, Elizabeth L; Jennings, Scott; Kraberger, Simona; Farkas, Kata; Julian, Laurel; Massaro, Melanie; Ballard, Grant; Ainley, David G

    2015-04-01

    Little is known about viruses associated with Antarctic animals, although they are probably widespread. We recovered a novel polyomavirus from Adélie penguin (Pygoscelis adeliae) faecal matter sampled in a subcolony at Cape Royds, Ross Island, Antarctica. The 4988 nt Adélie penguin polyomavirus (AdPyV) has a typical polyomavirus genome organization with three ORFs that encoded capsid proteins on the one strand and two non-structural protein-coding ORFs on the complementary strand. The genome of AdPyV shared ~60 % pairwise identity with all avipolyomaviruses. Maximum-likelihood phylogenetic analysis of the large T-antigen (T-Ag) amino acid sequences showed that the T-Ag of AdPyV clustered with those of avipolyomaviruses, sharing between 48 and 52 % identities. Only three viruses associated with Adélie penguins have been identified at a genomic level, avian influenza virus subtype H11N2 from the Antarctic Peninsula and, respectively, Pygoscelis adeliae papillomavirus and AdPyV from capes Crozier and Royds on Ross Island.

  18. The dynamics of herpesvirus and polyomavirus reactivation and shedding in healthy adults: a 14-month longitudinal study

    NASA Technical Reports Server (NTRS)

    Ling, Paul D.; Lednicky, John A.; Keitel, Wendy A.; Poston, David G.; White, Zoe S.; Peng, RongSheng; Liu, Zhensheng; Mehta, Satish K.; Pierson, Duane L.; Rooney, Cliona M.; Vilchez, Regis A.; Smith, E. O'Brian; Butel, Janet S.

    2003-01-01

    Humans are infected with viruses that establish long-term persistent infections. To address whether immunocompetent individuals control virus reactivation globally or independently and to identify patterns of sporadic reactivation, we monitored herpesviruses and polyomaviruses in 30 adults, over 14 months. Epstein-Barr virus (EBV) DNA was quantitated in saliva and peripheral blood mononuclear cells (PBMCs), cytomegalovirus (CMV) was assayed in urine, and JC virus (JCV) and BK virus (BKV) DNAs were assayed in urine and PBMCs. All individuals shed EBV in saliva, whereas 67% had >or=1 blood sample positive for EBV. Levels of EBV varied widely. CMV shedding occurred infrequently but occurred more commonly in younger individuals (P<.03). JCV and BKV virurias were 46.7% and 0%, respectively. JCV shedding was age dependent and occurred commonly in individuals >or=40 years old (P<.03). Seasonal variation was observed in shedding of EBV and JCV, but there was no correlation among shedding of EBV, CMV, and JCV (P>.50). Thus, adults independently control persistent viruses, which display discordant, sporadic reactivations.

  19. No evidence of Polyomavirus and EBV infections in Italian patients with mixed cryoglobulinemia infected chronically with HCV.

    PubMed

    Comar, Manola; Zanotta, Nunzia; Del Savio, Rossella; Vascotto, Fulvia; Calabrese, Nadia; Zorat, Francesca; Pozzato, Gabriele

    2014-04-01

    Mixed cryoglobulinemia is a lymphoproliferative disorder associated with hepatitis C virus (HCV). In patients chronically affected by HCV the prevalence of mixed cryoglobulinemia is variable ranging from 0% to 56%. To verify whether polyomaviruses (PyV) play a role in this disorder a total of 222 blood samples from 63 HCV chronic patients, 43 with mixed cryoglobulinemia, 59 chronic lymphocytic leukemia, 50 polytransfused patients, and 50 blood donors were evaluated for Merkel (MCPyV), BKV, JCV, and SV40. EBV was additionally included in the analysis since association with this disorder has been reported. Mixed cryoglobulinemia patients infected chronically with HCV resulted negative for both PyV and EBV. MCPyV was found in 1 subject with Merkel Cell Carcinoma, in 10% of polytransfused and in 10% of blood donors while EBV was detected in 22% of polytransfused, 10% of B-cell lymphatic leukemia patients and 4% of blood donors (P < 0.01). Taken together, the absence of PyV and EBV in HCV-mixed cryoglobulinemia patients seems to exclude a direct involvement of these viruses in the pathogenesis of this disease while the presence of MCPyV in healthy individuals, at the same rate as in polytransfused patients, may reinforce data on a minimal role of this virus in other human pathologies.

  20. Genetic Analysis Workshop 18: Methods and strategies for analyzing human sequence and phenotype data in members of extended pedigrees

    PubMed Central

    2014-01-01

    Genetic Analysis Workshop 18 provided a platform for developing and evaluating statistical methods to analyze whole-genome sequence data from a pedigree-based sample. In this article we present an overview of the data sets and the contributions that analyzed these data. The family data, donated by the Type 2 Diabetes Genetic Exploration by Next-Generation Sequencing in Ethnic Samples Consortium, included sequence-level genotypes based on sequencing and imputation, genome-wide association genotypes from prior genotyping arrays, and phenotypes from longitudinal assessments. The contributions from individual research groups were extensively discussed before, during, and after the workshop in theme-based discussion groups before being submitted for publication. PMID:25519310

  1. COINFECTION OF CALIFORNIA SEA LION ADENOVIRUS 1 AND A NOVEL POLYOMAVIRUS IN A HAWAIIAN MONK SEAL (NEOMONACHUS SCHAUINSLANDI).

    PubMed

    Cortés-Hinojosa, Galaxia; Doescher, Bethany; Kinsel, Michael; Lednicky, John; Loeb, Julia; Waltzek, Thomas; Wellehan, James F X

    2016-06-01

    The Hawaiian monk seal (Neomonachus schauinslandi) is an endangered species. Here, we present a clinical case of a 26-yr-old male Hawaiian monk seal (HMS) kept in an aquarium with a history of intermittent anorexia and evidence of renal disease. Histologic examination revealed eosinophilic intranuclear inclusions in the liver. Conventional nested PCR protocols were used to test for viruses, and it tested positive for adenovirus and polyomavirus, and negative for herpesvirus. The adenovirus partial polymerase gene is 100% homologous to that of California sea lion adenovirus 1 (CSLAdV-1). CSLAdV-1 causes viral hepatitis in CSL, and has recently been reported in different species of otariids in an aquarium in Japan ( Otaria flavescens and Arctocephalus pusillus ) and a sequence from Spain has been submitted in NCBI as Otaria flavescens adenovirus-1. The polyomavirus in this animal is a novel virus, and is the first polyomavirus discovered in Hawaiian monk seals. This new virus is designated Hawaiian monk seal polyomavirus (HMSPyV-1), and is 83% homologous to California sea lion Polyomavirus-1 (CSLPyV-1). This is the first report of viral coinfection in a HMS and clinical significance in this case remains unclear but may be associated with advanced age.

  2. Creating Fantastic PI Workshops

    SciTech Connect

    Biedermann, Laura B.; Clark, Blythe G.; Colbert, Rachel S.; Dagel, Amber Lynn; Gupta, Vipin P.; Hibbs, Michael R.; Perkins, David Nikolaus; West, Roger Derek

    2015-10-01

    The goal of this SAND report is to provide guidance for other groups hosting workshops and peerto-peer learning events at Sandia. Thus this SAND report provides detail about our team structure, how we brainstormed workshop topics and developed the workshop structure. A Workshop “Nuts and Bolts” section provides our timeline and check-list for workshop activities. The survey section provides examples of the questions we asked and how we adapted the workshop in response to the feedback.

  3. 78 FR 12763 - Fecal Microbiota for Transplantation; Public Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-25

    ... HUMAN SERVICES Food and Drug Administration Fecal Microbiota for Transplantation; Public Workshop AGENCY... ``Fecal Microbiota for Transplantation.'' The purpose of the public workshop is to exchange information... fecal microbiota for transplantation (FMT). ] Date and Time: The public workshop will be held on May...

  4. 76 FR 37118 - Manual Materials Handling (MMH) Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-24

    ... HUMAN SERVICES Centers for Disease Control and Prevention Manual Materials Handling (MMH) Workshop... Health, will be holding a two-day Manual Materials Handling (MMH) Workshop. The Workshop is a National... engineering solutions for manual materials handling jobs in Retail, Wholesale and Warehouse industries....

  5. BK polyomavirus with archetypal and rearranged non-coding control regions is present in cerebrospinal fluids from patients with neurological complications.

    PubMed

    Bárcena-Panero, Ana; Echevarría, Juan E; Van Ghelue, Marijke; Fedele, Giovanni; Royuela, Enrique; Gerits, Nancy; Moens, Ugo

    2012-08-01

    BK polyomavirus (BKPyV) has recently been postulated as an emerging opportunistic pathogen of the human central nervous system (CNS), but it is not known whether specific strains are associated with the neurotropic character of BKPyV. The presence of BKPyV large T-antigen DNA was examined in 2406 cerebrospinal fluid (CSF) samples from neurological patients with suspected JC polyomavirus infection. Twenty patients had a large T-antigen DNA-positive specimen. The non-coding control region (NCCR) of the BKPyV strains amplified from CSF from these 20 patients, strains circulating in renal and bone marrow transplant recipients and from healthy pregnant women was sequenced. The archetypal conformation was the most prevalent in all groups and 14 of the neurological patients harboured archetypal strains, while the remaining six patients possessed BKPyV with rearranged NCCR similar to previously reported variants from non-neurological patients. Transfection studies in Vero cells revealed that five of six early and four of six late rearranged promoters of these CSF isolates showed significantly higher activity than the corresponding archetypal promoter. From seven of the neurological patients with BKPyV DNA-positive CSF, paired serum samples were available. Five of them were negative for BKPyV DNA, while serum from the remaining two patients harboured BKPyV strains with archetypal NCCR that differed from those present in their CSF. Our results suggest that NCCR rearrangements are not a hallmark for BKPyV neurotropism and the dissemination of a rearranged NCCR from the blood may not be the origin of BKPyV CNS infection.

  6. Mutations in the putative calcium-binding domain of polyomavirus VP1 affect capsid assembly

    NASA Technical Reports Server (NTRS)

    Haynes, J. I. 2nd; Chang, D.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    Calcium ions appear to play a major role in maintaining the structural integrity of the polyomavirus and are likely involved in the processes of viral uncoating and assembly. Previous studies demonstrated that a VP1 fragment extending from Pro-232 to Asp-364 has calcium-binding capabilities. This fragment contains an amino acid stretch from Asp-266 to Glu-277 which is quite similar in sequence to the amino acids that make up the calcium-binding EF hand structures found in many proteins. To assess the contribution of this domain to polyomavirus structural integrity, the effects of mutations in this region were examined by transfecting mutated viral DNA into susceptible cells. Immunofluorescence studies indicated that although viral protein synthesis occurred normally, infective viral progeny were not produced in cells transfected with polyomavirus genomes encoding either a VP1 molecule lacking amino acids Thr-262 through Gly-276 or a VP1 molecule containing a mutation of Asp-266 to Ala. VP1 molecules containing the deletion mutation were unable to bind 45Ca in an in vitro assay. Upon expression in Escherichia coli and purification by immunoaffinity chromatography, wild-type VP1 was isolated as pentameric, capsomere-like structures which could be induced to form capsid-like structures upon addition of CaCl2, consistent with previous studies. However, although VP1 containing the point mutation was isolated as pentamers which were indistinguishable from wild-type VP1 pentamers, addition of CaCl2 did not result in their assembly into capsid-like structures. Immunogold labeling and electron microscopy studies of transfected mammalian cells provided in vivo evidence that a mutation in this region affects the process of viral assembly.

  7. Dynamics of pregnancy-associated polyomavirus urinary excretion: a prospective longitudinal study.

    PubMed

    McClure, Gloria B; Gardner, J Suzette; Williams, Jason T; Copeland, Christina M; Sylvester, Sarah K; Garcea, Robert L; Meinerz, Natalie M; Groome, Lynn J; Vanchiere, John A

    2012-08-01

    Asymptomatic polyomaviruria of pregnancy has been documented in point prevalence studies, but little attention has been given to the dynamics of polyomavirus excretion during pregnancy because of its benign course. We tested the hypothesis that the frequency and/or magnitude of polyomavirus excretion would increase as pregnancy progresses. Urine specimens were obtained prospectively from 179 healthy women during uncomplicated pregnancies and 37 healthy non-pregnant women. Real-time polymerase chain reaction was used to determine BK virus (BKV) and JC virus (JCV) viral loads in urine, blood, and rectal and vaginal swabs collected during routine obstetric and gynecologic clinic visits. Asymptomatic urinary shedding of BKV and/or JCV was observed in 384 (48.0%) of 800 specimens from 100 (55.8%) pregnant women. BKV excretion was more common in pregnant than non-pregnant women (41.3% vs. 13.5%, P = 0.0026). The frequency of JCV excretion was no different in pregnant compared to non-pregnant women. The frequency and magnitude of polyomavirus shedding did not vary with gestational age. Post-partum shedding of BKV, but not JCV, rapidly decreased to undetectable levels. Pregnancy-associated BKV excretion begins early in pregnancy and terminates rapidly post-partum. Neither the frequency nor magnitude of BKV or JCV shedding increased with pregnancy progression. Further study into the host factors that regulate pregnancy-associated BKV excretion may allow identification of the host factors that predict susceptibility to BKV-associated diseases in immune compromised patients.

  8. Isolation and characterization of NIH 3T3 cells expressing polyomavirus small T antigen

    SciTech Connect

    Noda, T.; Satake, M.; Robins, T.; Ito, Y.

    1986-10-01

    The polyomavirus small T-antigen gene, together with the polyomavirus promoter, was inserted into retrovirus vector pGV16 which contains the Moloney sarcoma virus long terminal repeat and neomycin resistance gene driven by the simian virus 40 promoter. This expression vector, pGVST, was packaged into retrovirus particles by transfection of PSI2 cells which harbor packaging-defective murine retrovirus genome. NIH 3T3 cells were infected by this replication-defective retrovirus containing pGVST. Of the 15 G418-resistant cell clones, 8 express small T antigen at various levels as revealed by immunoprecipitation. A cellular protein with an apparent molecular weight of about 32,000 coprecipitates with small T antigen. Immunofluorescent staining shows that small T antigen is mainly present in the nuclei. Morphologically, cells expressing small T antigen are indistinguishable from parental NIH 3T3 cells and have a microfilament pattern similar to that in parental NIH 3T3 cells. Cells expressing small T antigen form a flat monolayer but continue to grow beyond the saturation density observed for parental NIH 3T3 cells and eventually come off the culture plate as a result of overconfluency. There is some correlation between the level of expression of small T antigen and the growth rate of the cells. Small T-antigen-expressing cells form small colonies in soft agar. However, the proportion of cells which form these small colonies is rather small. A clone of these cells tested did not form tumors in nude mice within 3 months after inoculation of 10/sup 6/ cells per animal. Thus, present studies establish that the small T antigen of polyomavirus is a second nucleus-localized transforming gene product of the virus (the first one being large T antigen) and by itself has a function which is to stimulate the growth of NIH 3T3 cells beyond their saturation density in monolayer culture.

  9. Successful pregnancy in renal transplant recipient with previous known polyomavirus nephropathy.

    PubMed

    Midtvedt, Karsten; Bjorang, Ola; Letting, Anne-Sofie

    2007-01-01

    Pregnancy after renal transplantation has become increasingly common. Studies in non-immunocompromised patients have shown that pregnant women have increased susceptibility to infection or reactivation of latent virus such as BK virus. To what extent a renal transplant recipient is at risk for reactivation of polyoma virus during pregnancy remains unknown. We hereby report successful pregnancy outcome in a renal transplant recipient with a known history of BK virus nephropathy treated with cidofovir i.v. To our knowledge, this is the first published experience with a successful pregnancy in renal transplant recipients with known history of polyomavirus-associated nephropathy.

  10. Identification of a UV-induced trans-acting protein that stimulates polyomavirus DNA replication

    SciTech Connect

    Ronai, Z.A.; Weinstein, I.B. )

    1988-03-01

    Previous studies provided indirect evidence that the ability of a variety of DNA-damaging agents to induce asynchronous polyomavirus DNA replication in the H3 rat fibroblast cell line is mediated by a trans-acting factor. Using an erythrocyte insertion technique to introduce protein fractions from UV-irradiated cells into unirradiated H3 cells, we have now obtained evidence that this factor is a 60-kilodalton protein. These findings provide evidence that DNA damage in mammalian cells induces a factor that can alter the replication of a viral DNA.

  11. Interobserver agreement for Polyomavirus nephropathy grading in renal allografts using the working proposal from the 10th Banff Conference on Allograft Pathology.

    PubMed

    Sar, Aylin; Worawichawong, Suchin; Benediktsson, Hallgrimur; Zhang, Jianguo; Yilmaz, Serdar; Trpkov, Kiril

    2011-12-01

    A classification schema for grading Polyomavirus nephropathy was proposed at the 2009 Banff allograft meeting. The schema included 3 stages of Polyomavirus nephropathy: early (stage A), florid (stage B), and late sclerosing (stage C). Grading categories for histologic viral load levels were also proposed. To examine the applicability and the interobserver agreement of the proposed Polyomavirus nephropathy grading schema, we evaluated 24 renal allograft biopsies with confirmed Polyomavirus nephropathy by histology and SV40. Four renal pathologists independently scored the Polyomavirus nephropathy stage (A, B, or C), without knowledge of the clinical history. Viral load was scored as a percent of tubules exhibiting viral replication, using either a 3-tier viral load score (1: ≤1%; 2: >1%-10%; 3: >10%) or a 4-tier score (1: ≤1%; 2: >1%-≤5%; 3: >5%-15%; 4: >15%). The κ score for the Polyomavirus nephropathy stage was 0.47 (95% confidence interval, 0.35-0.60; P < .001). There was a substantial agreement using both the 3-tier and the 4-tier scoring for the viral load (Kendall concordance coefficients, 0.72 and 0.76, respectively; P < .001 for both). A better complete agreement was found using the 3-tier viral load score. In this first attempt to evaluate the interobserver reproducibility of the proposed Polyomavirus nephropathy classifying schema, we demonstrated moderate κ agreement in assessing the Polyomavirus nephropathy stage and a substantial agreement in scoring the viral load level. The proposed grading schema can be applied in routine allograft biopsy practice for grading the Polyomavirus nephropathy stage and the viral load level.

  12. Craniofacial morphogenesis workshop report.

    PubMed

    Solursh, M; Murray, J

    1994-05-01

    The following report highlights the discussions and interaction at the workshop on craniofacial morphogenesis, sponsored by The Human Frontier Science Program, held in April 1993 at the University of Iowa. A brief summary of selected sessions is included to exemplify the benefits of bringing together individuals from various disciplines and backgrounds in order to establish a unified theory of craniofacial morphogenesis. The synthesis of information and experience of a wide range of approaches made the 4-day period an invaluable experience for the participants from nine different countries.

  13. Social Technologies in Shipbuilding Workshop

    DTIC Science & Technology

    1983-05-01

    oriented ( behavior modification ) approaches. Process discussions covered topics ranging from preconditions to undertaking a change project, to start...quality circles, quality of work life, 2) work redesign or sociotechnical systems (autonomous work groups, multi-skilled workers), and 3) behavior ... modification (human performance engineering, performance management). The overall objective of the workshop was to examine both the process and content

  14. Report from the Fifth National Cancer Institute Mouse Models of Human Cancers Consortium Nervous System Tumors Workshop

    PubMed Central

    Gutmann, David H.; Stiles, Charles D.; Lowe, Scott W.; Bollag, Gideon E.; Furnari, Frank B.; Charest, Al

    2011-01-01

    Cancers of the nervous system are clinically challenging tumors that present with varied histopathologies and genetic etiologies. While the prognosis for the most malignant of these tumors is essentially unchanged despite decades of basic and translational science research, the past few years have witnessed the identification of numerous targetable molecular alterations in these cancers. With the advent of advanced genomic sequencing methodologies and the development of accurate small-animal models of these nervous system cancers, we are now ideally positioned to develop personalized therapies that target the unique cellular and molecular changes that define their formation and drive their continued growth. Recently, the National Cancer Institute convened a workshop to advance our understanding of nervous system cancer mouse models and to inform clinical trials by reconsidering these neoplasms as complex biological systems characterized by heterogeneity at all levels. PMID:21727208

  15. Workshop introduction

    SciTech Connect

    Streeper, Charles

    2010-01-01

    The Department of Energy's National Nuclear Security Administration's Global Threat Reduction Initiative (GTRI) has three subprograms that directly reduce the nuclear/radiological threat; Convert (Highly Enriched Uranium), Protect (Facilities), and Remove (Materials). The primary mission of the Off-Site Source Recovery Project (OSRP) falls under the 'Remove' subset. The purpose of this workshop is to provide a venue for joint-technical collaboration between the OSRP and the Nuclear Radiation Safety Service (NRSS). Eisenhower's Atoms for Peace initiative and the Soviet equivalent both promoted the spread of the paradoxical (peaceful and harmful) properties of the atom. The focus of nonproliferation efforts has been rightly dedicated to fissile materials and the threat they pose. Continued emphasis on radioactive materials must also be encouraged. An unquantifiable threat still exists in the prolific quantity of sealed radioactive sources (sources) spread worldwide. It does not appear that the momentum of the evolution in the numerous beneficial applications of radioactive sources will subside in the near future. Numerous expert studies have demonstrated the potentially devastating economic and psychological impacts of terrorist use of a radiological dispersal or emitting device. The development of such a weapon, from the acquisition of the material to the technical knowledge needed to develop and use it, is straightforward. There are many documented accounts worldwide of accidental and purposeful diversions of radioactive materials from regulatory control. The burden of securing sealed sources often falls upon the source owner, who may not have a disposal pathway once the source reaches the end of its useful life. This disposal problem is exacerbated by some source owners not having the resources to safely and compliantly store them. US Nuclear Regulatory Commission (NRC) data suggests that, in the US alone, there are tens of thousands of high-activity (IAEA

  16. Rapid detection of urinary polyomavirus BK by heterodyne-based surface plasmon resonance biosensor

    NASA Astrophysics Data System (ADS)

    Su, Li-Chen; Tian, Ya-Chung; Chang, Ying-Feng; Chou, Chien; Lai, Chao-Sung

    2014-01-01

    In renal transplant patients, immunosuppressive therapy may result in the reactivation of polyomavirus BK (BKV), leading to polyomavirus-associated nephropathy (PVAN), which inevitably causes allograft failure. Since the treatment outcomes of PVAN remain unsatisfactory, early identification and continuous monitoring of BKV reactivation and reduction of immunosuppressants are essential to prevent PVAN development. The present study demonstrated that the developed dual-channel heterodyne-based surface plasmon resonance (SPR) biosensor is applicable for the rapid detection of urinary BKV. The use of a symmetrical reference channel integrated with the poly(ethylene glycol)-based low-fouling self-assembled monolayer to reduce the environmental variations and the nonspecific noise was proven to enhance the sensitivity in urinary BKV detection. Experimentally, the detection limit of the biosensor for BKV detection was estimated to be around 8500 copies/mL. In addition, urine samples from five renal transplant patients were tested to rapidly distinguish PVAN-positive and PVAN-negative renal transplant patients. By virtue of its simplicity, rapidity, and applicability, the SPR biosensor is a remarkable potential to be used for continuous clinical monitoring of BKV reactivation.

  17. Polyomavirus DNA is damaged in target cells during cytotoxic T-lymphocyte-mediated killing

    SciTech Connect

    Sellins, K.S.; Cohen, J.J.

    1989-02-01

    Target cell DNA damage is an early event in cytotoxic T-lymphocyte (CTS)-mediated killing. It has been hypothesized that this DNA damage may serve as one mechanism of destroying viral genetic material inside infected cells. The authors directly examined the fate of viral DNA in target cells during CTL-mediated lysis. Polyomavirus DNA in transfected murine P815 mastocytoma targets was digested along with cellular DNA into oligonucleosome-sized fragments, although intact forms, possibly virion-associated DNA, were also present. In infected BALB/3T3 murine fibroblasts, which normally undergo single-stranded nicks when killed by CTL, polyomavirus DNA was converted to relaxed forms in the presence of CTL. These results suggest that the fate of the viral DNA depends on the stage of the viral life cycle and corresponds to the fate of the host cell DNA. Cleavage of the viral genome prior to assembly may thus be an important mechanism in specific antiviral immunity.

  18. 77 FR 43846 - Food and Drug Administration Pediatric Medical Devices Workshop; Notice of Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-26

    ... HUMAN SERVICES Food and Drug Administration Food and Drug Administration Pediatric Medical Devices... Administration's (FDA) Office of Orphan Products Development is announcing the following workshop: FDA Pediatric Medical Devices Workshop. This meeting is intended to focus on challenges in pediatric device...

  19. Merkel Cell Polyomavirus Small T Antigen Induces Cancer and Embryonic Merkel Cell Proliferation in a Transgenic Mouse Model

    PubMed Central

    Geng, Xuehui; Shuda, Yoko; Ostrowski, Stephen M.; Lukianov, Stefan; Jenkins, Frank J.; Honda, Kord; Maricich, Stephen M.; Moore, Patrick S.; Chang, Yuan

    2015-01-01

    Merkel cell polyomavirus (MCV) causes the majority of human Merkel cell carcinomas (MCC) and encodes a small T (sT) antigen that transforms immortalized rodent fibroblasts in vitro. To develop a mouse model for MCV sT-induced carcinogenesis, we generated transgenic mice with a flox-stop-flox MCV sT sequence homologously recombined at the ROSA locus (ROSAsT), allowing Cre-mediated, conditional MCV sT expression. Standard tamoxifen (TMX) administration to adult UbcCreERT2; ROSAsT mice, in which Cre is ubiquitously expressed, resulted in MCV sT expression in multiple organs that was uniformly lethal within 5 days. Conversely, most adult UbcCreERT2; ROSAsT mice survived low-dose tamoxifen administration but developed ear lobe dermal hyperkeratosis and hypergranulosis. Simultaneous MCV sT expression and conditional homozygous p53 deletion generated multi-focal, poorly-differentiated, highly anaplastic tumors in the spleens and livers of mice after 60 days of TMX treatment. Mouse embryonic fibroblasts from these mice induced to express MCV sT exhibited anchorage-independent cell growth. To examine Merkel cell pathology, MCV sT expression was also induced during mid-embryogenesis in Merkel cells of Atoh1CreERT2/+; ROSAsT mice, which lead to significantly increased Merkel cell numbers in touch domes at late embryonic ages that normalized postnatally. Tamoxifen administration to adult Atoh1CreERT2/+; ROSAsT and Atoh1CreERT2/+; ROSAsT; p53flox/flox mice had no effects on Merkel cell numbers and did not induce tumor formation. Taken together, these results show that MCV sT stimulates progenitor Merkel cell proliferation in embryonic mice and is a bona fide viral oncoprotein that induces full cancer cell transformation in the p53-null setting. PMID:26544690

  20. Junior Electronics Workshops and Their Questionnaires Reviews

    NASA Astrophysics Data System (ADS)

    Muto, Cosy; Maruta, Shuichiro; Noyori, Kazumasa; Yanaida, Masashi

    In this paper, a trial to educate electronics for both elementary pupils and junior-high students is reported. A “making your own radio” workshop for elementary kids features a paper-craft resonator made of toilet paper cores and an empty box of tissue papers as well as solder-less main radio circuit. For elder elementary and junior-high pupils, a workshop making a bat detector (an ultra-sonic receiver) is provided to help their summer vacation research. Both workshops are planned to enlarge students wishing to knock the door of electronics. Also, we report questionnaires results for those workshops and follow up research results for bat detector workshop. Those results show that both children and parents long for good experiences on science/electronics materials and these experiences are important for future human resources in scientific fields including analog electronics.

  1. Structural Based Analyses of the JC Virus T-Antigen F258L Mutant Provides Evidence for DNA Dependent Conformational Changes in the C-Termini of Polyomavirus Origin Binding Domains

    PubMed Central

    Meinke, Gretchen; Phelan, Paul J.; Shin, Jong; Gagnon, David; Archambault, Jacques; Bohm, Andrew; Bullock, Peter A.

    2016-01-01

    The replication of human polyomavirus JCV, which causes Progressive Multifocal Leukoencephalopathy, is initiated by the virally encoded T-antigen (T-ag). The structure of the JC virus T-ag origin-binding domain (OBD) was recently solved by X-ray crystallography. This structure revealed that the OBD contains a C-terminal pocket, and that residues from the multifunctional A1 and B2 motifs situated on a neighboring OBD molecule dock into the pocket. Related studies established that a mutation in a pocket residue (F258L) rendered JCV T-ag unable to support JCV DNA replication. To establish why this mutation inactivated JCV T-ag, we have solved the structure of the F258L JCV T-ag OBD mutant. Based on this structure, it is concluded that the structural consequences of the F258L mutation are limited to the pocket region. Further analyses, utilizing the available polyomavirus OBD structures, indicate that the F258 region is highly dynamic and that the relative positions of F258 are governed by DNA binding. The possible functional consequences of the DNA dependent rearrangements, including promotion of OBD cycling at the replication fork, are discussed. PMID:26735515

  2. Structural Based Analyses of the JC Virus T-Antigen F258L Mutant Provides Evidence for DNA Dependent Conformational Changes in the C-Termini of Polyomavirus Origin Binding Domains.

    PubMed

    Meinke, Gretchen; Phelan, Paul J; Shin, Jong; Gagnon, David; Archambault, Jacques; Bohm, Andrew; Bullock, Peter A

    2016-01-01

    The replication of human polyomavirus JCV, which causes Progressive Multifocal Leukoencephalopathy, is initiated by the virally encoded T-antigen (T-ag). The structure of the JC virus T-ag origin-binding domain (OBD) was recently solved by X-ray crystallography. This structure revealed that the OBD contains a C-terminal pocket, and that residues from the multifunctional A1 and B2 motifs situated on a neighboring OBD molecule dock into the pocket. Related studies established that a mutation in a pocket residue (F258L) rendered JCV T-ag unable to support JCV DNA replication. To establish why this mutation inactivated JCV T-ag, we have solved the structure of the F258L JCV T-ag OBD mutant. Based on this structure, it is concluded that the structural consequences of the F258L mutation are limited to the pocket region. Further analyses, utilizing the available polyomavirus OBD structures, indicate that the F258 region is highly dynamic and that the relative positions of F258 are governed by DNA binding. The possible functional consequences of the DNA dependent rearrangements, including promotion of OBD cycling at the replication fork, are discussed.

  3. Testing strategies for embryo-fetal toxicity of human pharmaceuticals. Animal models vs. in vitro approaches: a workshop report.

    PubMed

    van der Laan, Jan Willem; Chapin, Robert E; Haenen, Bert; Jacobs, Abigail C; Piersma, Aldert

    2012-06-01

    Reproductive toxicity testing is characterized by high animal use. For registration of pharmaceutical compounds, developmental toxicity studies are usually conducted in both rat and rabbits. Efforts have been underway for a long time to design alternatives to animal use. Implementation has lagged, partly because of uncertainties about the applicability domain of the alternatives. The reproductive cycle is complex and not all mechanisms of development can be mimicked in vitro. Therefore, efforts are underway to characterize the available alternative tests with regard to the mechanism of action they include. One alternative test is the mouse embryonic stem cell test (EST), which has been studied since the late 1990s. It is a genuine 3R "alternative" assay as it is essentially animal-free. A meeting was held to review the state-of-the-art of various in vitro models for prediction of developmental toxicity. Although the predictivity of individual assays is improving, a battery of several assays is likely to have even higher predictivity, which is necessary for regulatory acceptance. The workshop concluded that an important first step is a thorough survey of the existing rat and rabbit studies, to fully characterize the frequency of responses and the types of effects seen. At the same time, it is important to continue the optimization of in vitro assays. As more experience accumulates, the optimal conditions, assay structure, and applicability of the alternative assays are expected to emerge.

  4. Characterization of heavy metals and brominated flame retardants in the indoor and outdoor dust of e-waste workshops: implication for on-site human exposure.

    PubMed

    Xu, Feng; Liu, Yangcheng; Wang, Junxia; Zhang, Gang; Zhang, Wei; Liu, Lili; Wang, Jinfu; Pan, Bishu; Lin, Kuangfei

    2015-04-01

    Forty-four indoor and outdoor dust samples were collected from e-waste workshops and were analyzed to characterize the heavy metals and brominated flame retardants (BFRs) as well as on-site human exposure. The results showed that the most abundant Polybrominated diphenyl ethers (PBDE) congener from three sites was deca-BDE, and it was penta-BDE for the other site. A significant and positive association was found between BDE-209 and decabromodiphenyl ethane (DBDPE). The high percentage of nona-BDE indicated the debromination of deca-BDE during e-waste recycling. The ratio comparison of BDE-47 to (BDE-100 + BDE-99) indicated that the outdoor dust went through more physiochemical processes. The enrichment factors for Cu and Pb were high in both the indoor and outdoor samples. Cd significantly exceeded the Chinese soil guideline grade III. The PCA results combined with the enrichment factor (EF) values suggested common sources and behaviours of Cu, Pb and Sb in the indoor dust. Co, Cr, Ni, Zn and Mn in the outdoor samples were more likely affected by crust. Strong correlations were found only for Pb and Sb with polybrominated diphenyl ethers (PBDEs). The hazard index for on-site human exposure to Pb was at a chronic risk. Despite the low deleterious risk of BFRs, concern should be given to DBDPE; the chronic toxicity of which is not known.

  5. IPHE Infrastructure Workshop Proceedings

    SciTech Connect

    2010-02-01

    This proceedings contains information from the IPHE Infrastructure Workshop, a two-day interactive workshop held on February 25-26, 2010, to explore the market implementation needs for hydrogen fueling station development.

  6. Formaldehyde Workshop Agenda

    EPA Pesticide Factsheets

    This is the agenda for the Formaldehyde Workshop hosted by the Office of Research and Development's National Center for Environmental Assessments in cooperation with the IRIS Program. The workshop was held in April 2014

  7. Mars exploration study workshop 2

    NASA Technical Reports Server (NTRS)

    Duke, Michael B.; Budden, Nancy Ann

    1993-01-01

    A year-long NASA-wide study effort has led to the development of an innovative strategy for the human exploration of Mars. The latest Mars Exploration Study Workshop 2 advanced a design reference mission (DRM) that significantly reduces the perceived high costs, complex infrastructure, and long schedules associated with previous Mars scenarios. This surface-oriented philosophy emphasizes the development of high-leveraging surface technologies in lieu of concentrating exclusively on space transportation technologies and development strategies. As a result of the DRM's balanced approach to mission and crew risk, element commonality, and technology development, human missions to Mars can be accomplished without the need for complex assembly operations in low-Earth orbit. This report, which summarizes the Mars Exploration Study Workshop held at the Ames Research Center on May 24-25, 1993, provides an overview of the status of the Mars Exploration Study, material presented at the workshop, and discussions of open items being addressed by the study team. The workshop assembled three teams of experts to discuss cost, dual-use technology, and international involvement, and to generate a working group white paper addressing these issues. The three position papers which were generated are included in section three of this publication.

  8. Reading Workshop Survival Kit.

    ERIC Educational Resources Information Center

    Muschla, Gary Robert

    Intended for reading and classroom teachers, this book, organized in two parts, is a complete, step-by-step guide to setting up and running a reading workshop for grades 5-12 where reading is "the" priority. Part 1, "Management of the Reading Workshop," shows how to create a reading workshop, offers specific tools and…

  9. Thematic Issue: Workshops.

    ERIC Educational Resources Information Center

    Kirby, Michael, Ed.

    1978-01-01

    The articles in this publication trace the historical development of the theatre workshop, explain the relationship between the workshop and experimental theatre, and analyze the ways in which current drama workshops teach and develop the dramatic skills of the participants. The topics discussed include the special skills, production-oriented, and…

  10. ICP-MS Workshop

    SciTech Connect

    Carman, April J.; Eiden, Gregory C.

    2014-11-01

    This is a short document that explains the materials that will be transmitted to LLNL and DNN HQ regarding the ICP-MS Workshop held at PNNL June 17-19th. The goal of the information is to pass on to LLNL information regarding the planning and preparations for the Workshop at PNNL in preparation of the SIMS workshop at LLNL.

  11. Transformation by Polyomavirus Middle T Antigen Involves a Unique Bimodal Interaction with the Hippo Effector YAP

    PubMed Central

    Rouleau, Cecile; Pores Fernando, Arun T.; Hwang, Justin H.; Faure, Nathalie; Jiang, Tao; White, Elizabeth A.

    2016-01-01

    ABSTRACT Murine polyomavirus has repeatedly provided insights into tumorigenesis, revealing key control mechanisms such as tyrosine phosphorylation and phosphoinositide 3-kinase (PI3K) signaling. We recently demonstrated that polyomavirus small T antigen (ST) binds YAP, a major effector of Hippo signaling, to regulate differentiation. Here we characterize YAP as a target of middle T antigen (MT) important for transformation. Through a surface including residues R103 and D182, wild-type MT binds to the YAP WW domains. Mutation of either R103 or D182 of MT abrogates YAP binding without affecting binding to other signaling molecules or the strength of PI3K or Ras signaling. Either genetic abrogation of YAP binding to MT or silencing of YAP via short hairpin RNA (shRNA) reduced MT transformation, suggesting that YAP makes a positive contribution to the transformed phenotype. MT targets YAP both by activating signaling pathways that affect it and by binding to it. MT signaling, whether from wild-type MT or the YAP-binding MT mutant, promoted YAP phosphorylation at S127 and S381/397 (YAP2/YAP1). Consistent with the known functions of these phosphorylated serines, MT signaling leads to the loss of YAP from the nucleus and degradation. Binding of YAP to MT brings it together with protein phosphatase 2A (PP2A), leading to the dephosphorylation of YAP in the MT complex. It also leads to the enrichment of YAP in membranes. Taken together, these results indicate that YAP promotes MT transformation via mechanisms that may depart from YAP's canonical oncogenic transcriptional activation functions. IMPORTANCE The highly conserved Hippo/YAP pathway is important for tissue development and homeostasis. Increasingly, changes in this pathway are being associated with cancer. Middle T antigen (MT) is the primary polyomavirus oncogene responsible for tumor formation. In this study, we show that MT signaling promotes YAP phosphorylation, loss from the nucleus, and increased turnover

  12. TECHNICAL WORKSHOP ON HUMAN MILK SURVEILLANCE AND RESEARCH ON ENVIRONMENTAL CHEMICALS IN THE U.S.: AN OVERVIEW

    EPA Science Inventory

    Interest in human milk research and monitoring for environmental chemicals is growing, and as studies of chemicals in human milk are initiated, it is of the utmost importance that these studies be conducted using harmonized methods. Due to numerous limitations in previous studies...

  13. Goose Hemorrhagic polyomavirus detection in geese using real-time PCR assay.

    PubMed

    Leon, Olivier; Corrand, Léni; Bich, Tran Ngoc; Le Minor, Odile; Lemaire, Mylène; Guérin, Jean-Luc

    2013-12-01

    Goose hemorrhagic polyomavirus (GHPV) is the viral agent of hemorrhagic nephritis enteritis of geese (HNEG), a lethal disease of goslings. Although death is the most common outcome, geese that recover from HNEG are persistently infected. Here, we present the development of real-time SYBR Green real-time PCR targeted to GHPV and its use to assess the prevalence of GHPV infection in French geese flocks. When compared with classical end-point PCR, real-time PCR revealed a much better sensitivity and equivalent specificity. Real-time PCR could, therefore, be considered a gold standard for the detection of GHPV. Results of field investigations evidenced a very high prevalence of GHPV infections in French geese, largely associated with healthy carriage.

  14. Infectious Offspring: How Birds Acquire and Transmit an Avian Polyomavirus in the Wild

    PubMed Central

    Potti, Jaime; Blanco, Guillermo; Lemus, Jesús Á.; Canal, David

    2007-01-01

    Detailed patterns of primary virus acquisition and subsequent dispersal in wild vertebrate populations are virtually absent. We show that nestlings of a songbird acquire polyomavirus infections from larval blowflies, common nest ectoparasites of cavity-nesting birds, while breeding adults acquire and renew the same viral infections via cloacal shedding from their offspring. Infections by these DNA viruses, known potential pathogens producing disease in some bird species, therefore follow an ‘upwards vertical’ route of an environmental nature mimicking horizontal transmission within families, as evidenced by patterns of viral infection in adults and young of experimental, cross-fostered offspring. This previously undescribed route of viral transmission from ectoparasites to offspring to parent hosts may be a common mechanism of virus dispersal in many taxa that display parental care. PMID:18060070

  15. Whole-Genome Characterization and Genotyping of Global WU Polyomavirus Strains▿ †

    PubMed Central

    Bialasiewicz, Seweryn; Rockett, Rebecca; Whiley, David W.; Abed, Yacine; Allander, Tobias; Binks, Michael; Boivin, Guy; Cheng, Allen C.; Chung, Ju-Young; Ferguson, Patricia E.; Gilroy, Nicole M.; Leach, Amanda J.; Lindau, Cecilia; Rossen, John W.; Sorrell, Tania C.; Nissen, Michael D.; Sloots, Theo P.

    2010-01-01

    Exploration of the genetic diversity of WU polyomavirus (WUV) has been limited in terms of the specimen numbers and particularly the sizes of the genomic fragments analyzed. Using whole-genome sequencing of 48 WUV strains collected in four continents over a 5-year period and 16 publicly available whole-genome sequences, we identified three main WUV clades and five subtypes, provisionally termed Ia, Ib, Ic, II, IIIa, and IIIb. Overall nucleotide variation was low (0 to 1.2%). The discriminatory power of the previous VP2 fragment typing method was found to be limited, and a new, larger genotyping region within the VP2/1 interface was proposed. PMID:20357093

  16. Identification of amino acid sequences in the polyomavirus capsid proteins that serve as nuclear localization signals

    NASA Technical Reports Server (NTRS)

    Chang, D.; Haynes, J. I. Jr; Brady, J. N.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    The molecular mechanism participating in the transport of newly synthesized proteins from the cytoplasm to the nucleus in mammalian cells is poorly understood. Recently, the nuclear localization signal sequences (NLS) of many nuclear proteins have been identified, and most have been found to be composed of a highly basic amino acid stretch. A genetic "subtractive" and a biochemical "additive" approach were used in our studies to identify the NLS's of the polyomavirus structural capsid proteins. An NLS was identified at the N-terminus (Ala1-Pro-Lys-Arg-Lys-Ser-Gly-Val-Ser-Lys-Cys11) of the major capsid protein VP1 and at the C-terminus (Glu307 -Glu-Asp-Gly-Pro-Glu-Lys-Lys-Lys-Arg-Arg-Leu318) of the VP2/VP3 minor capsid proteins.

  17. The encapsidation of polyomavirus is not defined by a sequence-specific encapsidation signal.

    PubMed

    Spanielová, Hana; Fraiberk, Martin; Suchanová, Jiřina; Soukup, Jakub; Forstová, Jitka

    2014-02-01

    Mouse polyomavirus (MPyV) is considered a potential tool for the application of gene therapy; however, the current knowledge of the encapsulation of DNA into virions is vague. We used a series of assays based on the encapsidation of a reporter vector into MPyV pseudovirions to identify putative cis-acting elements that are involved in DNA encapsidation. None of the sequences that were derived from MPyV have been shown to solely enhance the encapsidation of a reporter vector in the assay. The frequency of encapsidation strongly correlated with the total intracellular amount of the vector after transfection. The encapsidation of target DNA into the pseudovirions was shown to be non-specific, and the packaging of non-replicated DNA was observed. We propose that the actual concentration of target DNA at the sites of virion formation is the primary factor that determines its selection for encapsidation.

  18. BK polyomavirus in kidney transplant recipients: screening, monitoring and clinical management.

    PubMed

    Varella, Rafael Brandão; Almeida, Jorge Reis; Lopes, Patrícia de Fátima; Matos, Jorge Paulo Strogoff de; Menezes, Paulo; Lugon, Jocemir Ronaldo

    2014-01-01

    BK polyomavirus (BKPyV) is a causal agent of nephropathy, ureteral stenosis and hemorrhagic cystitis in kidney transplant recipients, and is considered an important emerging disease in transplantation. Regular screening for BKPyV reactivation mainly during the first 2 years posttransplant, with subsequent pre-emptive reduction of immunosuppression is considered the best option to avoid disease progression, since successful clearance or reduction of viremia is achieved in the vast majority of patients within 6 months. The use of drugs with antiviral properties for patients with persistent viremia has been attempted despite unclear benefits. Clinical manifestations of BKPyV nephropathy, current strategies for diagnosis and monitoring of BKPyV infection, management of immunosuppressive regimen after detection of BKPyV reactivation and the use of antiviral drugs are discussed in this review.

  19. Identification of a nuclear localization sequence in the polyomavirus capsid protein VP2

    NASA Technical Reports Server (NTRS)

    Chang, D.; Haynes, J. I. 2nd; Brady, J. N.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1992-01-01

    A nuclear localization signal (NLS) has been identified in the C-terminal (Glu307-Glu-Asp-Gly-Pro-Gln-Lys-Lys-Lys-Arg-Arg-Leu318) amino acid sequence of the polyomavirus minor capsid protein VP2. The importance of this amino acid sequence for nuclear transport of newly synthesized VP2 was demonstrated by a genetic "subtractive" study using the constructs pSG5VP2 (expressing full-length VP2) and pSG5 delta 3VP2 (expressing truncated VP2, lacking amino acids Glu307-Leu318). These constructs were transfected into COS-7 cells, and the intracellular localization of the VP2 protein was determined by indirect immunofluorescence. These studies revealed that the full-length VP2 was localized in the nucleus, while the truncated VP2 protein was localized in the cytoplasm and not transported to the nucleus. A biochemical "additive" approach was also used to determine whether this sequence could target nonnuclear proteins to the nucleus. A synthetic peptide identical to VP2 amino acids Glu307-Leu318 was cross-linked to the nonnuclear proteins bovine serum albumin (BSA) or immunoglobulin G (IgG). The conjugates were then labeled with fluorescein isothiocyanate and microinjected into the cytoplasm of NIH 3T6 cells. Both conjugates localized in the nucleus of the microinjected cells, whereas unconjugated BSA and IgG remained in the cytoplasm. Taken together, these genetic subtractive and biochemical additive approaches have identified the C-terminal sequence of polyoma-virus VP2 (containing amino acids Glu307-Leu318) as the NLS of this protein.

  20. Polyomavirus middle-T antigen associates with the kinase domain of Src-related tyrosine kinases.

    PubMed Central

    Dunant, N M; Senften, M; Ballmer-Hofer, K

    1996-01-01

    Middle-T antigen of mouse polyomavirus, an oncogenic DNA virus, associates with and activates the cellular tyrosine kinases c-Src, c-Yes, and Fyn. This interaction is essential for polyomavirus-mediated transformation of cells in culture and tumor formation in animals. To determine the domain of c-Src directing association with middle-T, mutant c-Src proteins lacking the amino-terminal unique domain and the myristylation signal, the SH2 domain, the SH3 domain, or all three of these domains were coexpressed with middle-T in NIH 3T3 cells. All mutants were found to associate with middle-T, demonstrating that the kinase domain of c-Src, including the carboxy-terminal regulatory tail, is sufficient for association with middle-T. Moreover, we found that Hck, another member of the Src kinase family, does not bind middle-T, while chimeric kinases consisting of the amino-terminal domains of c-Src fused to the kinase domain of Hck or the amino-terminal domains of Hck fused to the kinase domain of c-Src associated with middle-T. Hck mutated at its carboxy-terminal regulatory residue, tyrosine 501, was also found to associate with middle-T. These results suggest that in Hck, the postulated intramolecular interaction between the carboxy-terminal regulatory tyrosine and the SH2 domain prevents association with middle-T. This intramolecular interaction apparently also limits the ability of c-Src to associate with middle-T, since removal of the SH2 or SH3 domain increases the efficiency with which middle-T binds c-Src. PMID:8627648

  1. Cellular proteins that associate with the middle and small T antigens of polyomavirus.

    PubMed

    Pallas, D C; Cherington, V; Morgan, W; DeAnda, J; Kaplan, D; Schaffhausen, B; Roberts, T M

    1988-11-01

    We have used two-dimensional gel electrophoresis to analyze in more detail the cellular proteins which associate with the middle and small tumor antigens (MT and ST, respectively) of polyomavirus. Proteins with molecular masses of 27, 29, 36, 51, 61, 63, and 85 kilodaltons (kDa) that specifically coimmunoprecipitated with MT were identified on these gels. The 36-, 51-, 61-, 63-, and 85-kDa proteins are probably the same as the proteins of similar sizes previously reported by a number of groups, whereas the 27- and 29-kDa proteins represent proteins that are heretofore undescribed. The 27- and 29-kDa proteins were abundant cellular proteins, whereas the others were minor cellular constituents. The association of each of these proteins with MT was sensitive to one or more mutations in MT that rendered it transformation defective. The association of the 85-kDa protein was the most sensitive indicator of the transformation competence of MT mutants. In addition, the 85-kDa protein was the only associated protein whose association with MT changed consistently in parallel with MT-associated phosphatidylinositol kinase activity. Furthermore, the fraction of the 85-kDa protein which was found associated with the MT complex contained 15 to 20% of its phosphate content on tyrosine. The 36- and 63-kDa proteins complexed with both polyomavirus MT and ST and comigrated on two-dimensional gels with two simian virus 40 ST-associated proteins originally described by Rundell and coworkers (K. Rundell, E. O. Major, and M. Lampert, J. Virol. 37:1090-1093, 1981). None of the other MT-associated proteins associated significantly with ST.

  2. Applied antineutrino physics workshop.

    SciTech Connect

    Lund, James C.

    2008-01-01

    This workshop is the fourth one of a series that includes the Neutrino Geophysics Conference at Honolulu, Hawaii, which I attended in 2005. This workshop was organized by the Astro-Particle and Cosmology laboratory in the recently opened Condoret building of the University of Paris. More information, including copies of the presentations, on the workshop is available on the website: www.apc.univ-paris7.fr/AAP2007/. The workshop aims at opening neutrino physics to various fields such that it can be applied in geosciences, nuclear industry (reactor and spent fuel monitoring) and non-proliferation. The workshop was attended by over 60 people from Europe, USA, Asia and Brazil. The meeting was also attended by representatives of the Comprehensive nuclear-Test Ban Treaty (CTBT) and the International Atomic Energy Agency (IAEA). The workshop also included a workshop dinner on board of a river boat sailing the Seine river.

  3. Report from the Light Water Reactor Sustainability Workshop on Advanced Instrumentation, Information, and Control Systems and Human-System Interface Technologies

    SciTech Connect

    Bruce P. Hallbert; J. J. Persensky; Carol Smidts; Tunc Aldemir; Joseph Naser

    2009-08-01

    . As an initial step in accomplishing this effort, the Light Water Reactor Sustainability Workshop on Advanced Instrumentation, Information, and Control Systems and Human-System Interface Technologies was held March 20–21, 2009, in Columbus, Ohio, to enable industry stakeholders and researchers in identification of the nuclear industry’s needs in the areas of future I&C technologies and corresponding technology gaps and research capabilities. Approaches for collaboration to bridge or fill the technology gaps were presented and R&D activities and priorities recommended. This report documents the presentations and discussions of the workshop and is intended to serve as a basis for the plan under development to achieve the goals of the I&C research pathway.

  4. Strengthening the Paediatricians Project 1: The need, content and process of a workshop to address the Priority Mental Health Disorders of adolescence in countries with low human resource for health

    PubMed Central

    2010-01-01

    delegates with basic information on adolescent psychiatry prior to the workshop was suggested to make the workshop more valuable. Conclusions There is a need to expand training for paediatricians from various backgrounds in adolescent psychiatry to strengthen their clinical skills to address the PMHD at the primary-care level. The evaluation suggests that the design and collaborative approach evident in this programme have merit as a model for training paediatricians in adolescent psychiatry in countries with low human resource for health. PMID:20167076

  5. 76 FR 42715 - Quarantine Release Errors in Blood Establishments; Public Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-19

    ... HUMAN SERVICES Food and Drug Administration Quarantine Release Errors in Blood Establishments; Public... Administration (FDA) is announcing a public workshop entitled: ``Quarantine Release Errors in Blood... workshop will focus on the extent and characteristics of QREs in blood establishments and...

  6. 77 FR 14814 - Tobacco Product Analysis; Scientific Workshop; Request for Comments

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-13

    ... HUMAN SERVICES Food and Drug Administration Tobacco Product Analysis; Scientific Workshop; Request for.... The Food and Drug Administration (FDA), Center for Tobacco Products is announcing a scientific... analyses are reliable and accurate. This scientific workshop will focus on understanding how...

  7. 77 FR 66850 - Public Workshop on Burkholderia: Exploring Current Issues and Identifying Regulatory Science Gaps

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-07

    ... HUMAN SERVICES Food and Drug Administration Public Workshop on Burkholderia: Exploring Current Issues and Identifying Regulatory Science Gaps AGENCY: Food and Drug Administration, HHS. ACTION: Notice of public workshop. SUMMARY: The Food and Drug Administration (FDA) is announcing the following...

  8. 75 FR 63478 - 5th Annual PHEMCE Stakeholders Workshop and BARDA Industry Day

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-15

    ... HUMAN SERVICES Office of the Secretary 5th Annual PHEMCE Stakeholders Workshop and BARDA Industry Day... Emergency Medical Countermeasures Enterprise (PHEMCE) Stakeholders Workshop and BARDA Industry Day to be..., International Governments, Industry, Healthcare Providers, First Responders, Community-Based Organizations,...

  9. 76 FR 55068 - Mobile Medical Applications Draft Guidance; Public Workshop; Correction

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-06

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration Mobile Medical Applications Draft Guidance; Public Workshop... FR 50231). The document announced a public workshop entitled ``Mobile ] Medical Applications...

  10. Merkel Cell Polyomavirus Exhibits Dominant Control of the Tumor Genome and Transcriptome in Virus-Associated Merkel Cell Carcinoma

    PubMed Central

    Starrett, Gabriel J.; Marcelus, Christina; Cantalupo, Paul G.; Katz, Joshua P.; Cheng, Jingwei; Akagi, Keiko; Thakuria, Manisha; Rabinowits, Guilherme; Wang, Linda C.; Symer, David E.; Pipas, James M.

    2017-01-01

    ABSTRACT Merkel cell polyomavirus is the primary etiological agent of the aggressive skin cancer Merkel cell carcinoma (MCC). Recent studies have revealed that UV radiation is the primary mechanism for somatic mutagenesis in nonviral forms of MCC. Here, we analyze the whole transcriptomes and genomes of primary MCC tumors. Our study reveals that virus-associated tumors have minimally altered genomes compared to non-virus-associated tumors, which are dominated by UV-mediated mutations. Although virus-associated tumors contain relatively small mutation burdens, they exhibit a distinct mutation signature with observable transcriptionally biased kataegic events. In addition, viral integration sites overlap focal genome amplifications in virus-associated tumors, suggesting a potential mechanism for these events. Collectively, our studies indicate that Merkel cell polyomavirus is capable of hijacking cellular processes and driving tumorigenesis to the same severity as tens of thousands of somatic genome alterations. PMID:28049147

  11. GH safety workshop position paper: A critical appraisal of recombinant human GH therapy in children and adults

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recombinant human Growth Hormone (rhGH) has been in use for 30 years, and over that time its safety and efficacy in children and adults has been subject to considerable scrutiny. In 2001, a statement from the GH Research Society (GRS) concluded that 'for approved indications, GH is safe'; however, t...

  12. Immunohistochemistry for Merkel cell polyomavirus is highly specific but not sensitive for the diagnosis of Merkel cell carcinoma in the Australian population.

    PubMed

    Paik, Julie Y; Hall, Geoffrey; Clarkson, Adele; Lee, Lianne; Toon, Christopher; Colebatch, Andrew; Chou, Angela; Gill, Anthony J

    2011-10-01

    Recent studies have demonstrated a high frequency of detection of Merkel cell polyomavirus in Merkel cell carcinoma. However, most of these studies are from European or North American centers that have relatively low sun exposure and may have a higher incidence of virus-driven oncogenesis compared with the highly sun-exposed but predominantly fair-skinned Australian population. We performed immunohistochemistry for Merkel cell polyomavirus on 104 cases of Merkel cell carcinoma and 74 cases of noncutaneous small cell-undifferentiated carcinoma from 3 major Australian centers. Nineteen (18.3%) cases of Merkel cell carcinoma showed positive staining for Merkel cell polyomavirus versus 1 (1.3%) of small cell-undifferentiated carcinoma. All 15 cases (14.3%) of Merkel cell carcinoma with areas of mixed squamous differentiation showed negative staining. We found positive staining in only 3 (7.7%) of 39 Merkel cell carcinoma from the head and neck (the most sun-exposed area) versus 16 (24.6%) of 65 of tumors from other sites (P < .05). Our findings support the concept of a Merkel cell polyomavirus-driven and a non-Merkel cell polyomavirus-driven (primarily sun-dependent) pathway in Merkel cell carcinoma carcinogenesis, with the latter being significantly more frequent in Australia and in mixed squamous-Merkel cell carcinoma (which is also more frequent in Australia). Although immunohistochemistry for Merkel cell polyomavirus seems to be highly specific in all populations, the low incidence of Merkel cell polyomavirus-positive Merkel cell carcinoma in a highly sun-exposed population limits its diagnostic utility in this setting.

  13. Anatomy and histology of rodent and human major salivary glands: -overview of the Japan salivary gland society-sponsored workshop-.

    PubMed

    Amano, Osamu; Mizobe, Kenichi; Bando, Yasuhiko; Sakiyama, Koji

    2012-10-31

    MAJOR SALIVARY GLANDS OF BOTH HUMANS AND RODENTS CONSIST OF THREE PAIRS OF MACROSCOPIC GLANDS: parotid, submandibular, and sublingual. These glands secrete serous, mucous or mixed saliva via the proper main excretory ducts connecting the glandular bodies with the oral cavity. A series of discoveries about the salivary ducts in the 17th century by Niels Stensen (1638-1686), Thomas Wharton (1614-1673), and Caspar Bartholin (1655-1738) established the concept of exocrine secretion as well as salivary glands. Recent investigations have revealed the endocrine functions of parotin and a variety of cell growth factors produced by salivary glands.The present review aims to describe macroscopic findings on the major salivary glands of rodents and the microscopic differences between those of humans and rodents, which review should be of interest to those researchers studying salivary glands.

  14. Proceedings of Workshop 1, the Human Brainmap Database Held in San Antonio, Texas on November 29-December 1, 1992.

    DTIC Science & Technology

    1993-02-17

    Atlases ......................................... 9 6. Choice of Development Environment .............................. 9 7. Distributed Development...a user interacts with the database. The GUI contains a digital atlas of the human brain adapted from the atlas of Talairach et al. (1967...Anatomically driven searches can be initiated by simply "paging" through the atlas and "clicking" on the area of interest. Anatomical searches can also be

  15. Temporal and geographic clustering of polyomavirus-associated olfactory tumors in 10 free-ranging raccoons (Procyon lotor).

    PubMed

    Giannitti, F; Higgins, R J; Pesavento, P A; Dela Cruz, F; Clifford, D L; Piazza, M; Parker Struckhoff, A; Del Valle, L; Bollen, A W; Puschner, B; Kerr, E; Gelberg, H; Mete, A; McGraw, S; Woods, L W

    2014-07-01

    Reports of primary nervous system tumors in wild raccoons are extremely rare. Olfactory tumors were diagnosed postmortem in 9 free-ranging raccoons from 4 contiguous counties in California and 1 raccoon from Oregon within a 26-month period between 2010 and 2012. We describe the geographic and temporal features of these 10 cases, including the laboratory diagnostic investigations and the neuropathologic, immunohistochemical, and ultrastructural characteristics of these tumors in the affected animals. All 9 raccoons from California were found within a localized geographic region of the San Francisco Bay Area (within a 44.13-km radius). The tight temporal and geographic clustering and consistent anatomic location in the olfactory system of tumor types not previously described in raccoons (malignant peripheral nerve sheath tumors and undifferentiated sarcomas) strongly suggest either a common cause or a precipitating factor leading to induction or potentiation of neuro-oncogenesis and so prompted an extensive diagnostic investigation to explore possible oncogenic infectious and/or toxic causes. By a consensus polymerase chain reaction strategy, a novel, recently reported polyomavirus called raccoon polyomavirus was identified in all 10 tumors but not in the normal brain tissue from the affected animals, suggesting that the virus might play a role in neuro-oncogenesis. In addition, expression of the viral protein T antigen was detected in all tumors containing the viral sequences. We discuss the potential role of raccoon polyomavirus as an oncogenic virus.

  16. Modeling and Simulation for Safeguards and Nonproliferation Workshop

    SciTech Connect

    Gilligan, Kimberly V.; Kirk, Bernadette Lugue

    2015-01-01

    The Modeling and Simulation for Safeguards and Nonproliferation Workshop was held December 15–18, 2014, at Oak Ridge National Laboratory. This workshop was made possible by the Next Generation Safeguards Initiative Human Capital Development (NGSI HCD) Program. The idea of the workshop was to move beyond the tried-and-true boot camp training of nonproliferation concepts to spend several days on the unique perspective of applying modeling and simulation (M&S) solutions to safeguards challenges.

  17. Tandem mirror theory workshop

    SciTech Connect

    1981-05-01

    The workshop was divided into three sections which were constituted according to subject matter: RF Heating, MHD Equilibrium and Stability, and Transport and Microstability. An overview from Livermore's point of view was given at the beginning of each session. Each session was assigned a secretary to take notes. These notes have been used in preparing this report on the workshop. The report includes the activities, conclusions, and recommendations of the workshop.

  18. Lunar Commercialization Workshop

    NASA Technical Reports Server (NTRS)

    Martin, Gary L.

    2008-01-01

    This slide presentation describes the goals and rules of the workshop on Lunar Commercialization. The goal of the workshop is to explore the viability of using public-private partnerships to open the new space frontier. The bulk of the workshop was a team competition to create a innovative business plan for the commercialization of the moon. The public private partnership concept is reviewed, and the open architecture as an infrastructure for potential external cooperation. Some possible lunar commercialization elements are reviewed.

  19. Solar education project workshop

    SciTech Connect

    Smith, J.B.

    1980-10-31

    A summary of proceedings of the Solar Education Project Workshop is presented. The workshop had as its focus the dissemination of curriculum materials developed by the Solar Energy Project of the New York State Department of Education under the sponsorship of the US Department of Energy. It includes, in addition to presentations by speakers and workshop leaders, specific comments from participants regarding materials available and energy-related activities underway in their respective states and suggested strategies from them for ongoing dissemination efforts.

  20. CARE 3 User's Workshop

    NASA Technical Reports Server (NTRS)

    1988-01-01

    A user's workshop for CARE 3, a reliability assessment tool designed and developed especially for the evaluation of high reliability fault tolerant digital systems, was held at NASA Langley Research Center on October 6 to 7, 1987. The main purpose of the workshop was to assess the evolutionary status of CARE 3. The activities of the workshop are documented and papers are included by user's of CARE 3 and NASA. Features and limitations of CARE 3 and comparisons to other tools are presented. The conclusions to a workshop questionaire are also discussed.

  1. Thermal Barrier Coating Workshop

    NASA Technical Reports Server (NTRS)

    Brindley, W. J. (Compiler); Lee, W. Y. (Compiler); Goedjen, J. G. (Compiler); Dapkunas, S. J. (Compiler)

    1995-01-01

    This document contains the agenda and presentation abstracts for the Thermal Barrier Coating Workshop, sponsored by NASA, DOE, and NIST. The workshop covered thermal barrier coating (TBC) issues related to applications, processing, properties, and modeling. The intent of the workshop was to highlight the state of knowledge on TBC's and to identify critical gaps in knowledge that may hinder TBC use in advanced applications. The workshop goals were achieved through presentations by 22 speakers representing industry, academia, and government as well as through extensive discussion periods.

  2. Fermilab Cryogenic Workshop Report

    SciTech Connect

    Hassenzahl, W. V.

    1980-06-18

    A workshop to discuss recent pressing problems experienced in the operation of helium refrigerators at the national laboratories was proposed by DOE. Early in 1980 it was decided that the workshop should be held at the Fermi National Accelerator Laboratory (Fermilab). The reasoning behind the selection of Fermilab included the proposed initial tests of the Central Liquefier, the recently experienced problems with refrigeration systems at Fermilab, and the fact that a previous workshop had been held at the Brookhaven National Laboratory, which, at present, would be the other logical choice for the workshop.

  3. Alternate fusion fuels workshop

    SciTech Connect

    Not Available

    1981-06-01

    The workshop was organized to focus on a specific confinement scheme: the tokamak. The workshop was divided into two parts: systems and physics. The topics discussed in the systems session were narrowly focused on systems and engineering considerations in the tokamak geometry. The workshop participants reviewed the status of system studies, trade-offs between d-t and d-d based reactors and engineering problems associated with the design of a high-temperature, high-field reactor utilizing advanced fuels. In the physics session issues were discussed dealing with high-beta stability, synchrotron losses and transport in alternate fuel systems. The agenda for the workshop is attached.

  4. Two overlapping sequence motifs within the polyomavirus enhancer are independently the targets of stimulation by both the tumor promoter 12-O-tetradecanoylphorbol-3-acetate and the Ha-ras oncogene

    SciTech Connect

    Yamaguchi, Yyko; Satake, Masanobu; Ito, Yoshiaki

    1989-03-01

    A tumor-promoting phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), strongly stimulates the activity of polyomavirus enhancer in a human erythroleukemia cell line, K562. The target of stimulation was the previously defined A element (from nucleotides 5107 to 5130) of the enhancer. The authors found that within the A element, two partly overlapping sequence motifs (one from nucleotides 5107 to 5117, the other from nucleotides 5113 to 5121) were independently the targets of TPA stimulation. The former is homologous to the enhancer core sequence of the adenovirus type 5 E1A gene, and the latter shares the consensus AP-1-binding site. In addition, transiently expressed Ha-ras oncogene also stimulated these two subelements in K562 cells, as they reported for NIH 3T3 cells previously.

  5. Nuclear thermal propulsion workshop overview

    NASA Technical Reports Server (NTRS)

    Clark, John S.

    1991-01-01

    NASA is planning an Exploration Technology Program as part of the Space Exploration Initiative to return U.S. astronauts to the moon, conduct intensive robotic exploration of the moon and Mars, and to conduct a piloted mission to Mars by 2019. Nuclear Propulsion is one of the key technology thrust for the human mission to Mars. The workshop addresses NTP (Nuclear Thermal Rocket) technologies with purpose to: assess the state-of-the-art of nuclear propulsion concepts; assess the potential benefits of the concepts for the mission to Mars; identify critical, enabling technologies; lay-out (first order) technology development plans including facility requirements; and estimate the cost of developing these technologies to flight-ready status. The output from the workshop will serve as a data base for nuclear propulsion project planning.

  6. Fifth International Microgravity Combustion Workshop

    NASA Technical Reports Server (NTRS)

    Sacksteder, Kurt (Compiler)

    1999-01-01

    This conference proceedings document is a compilation of 120 papers presented orally or as poster displays to the Fifth International Microgravity Combustion Workshop held in Cleveland, Ohio on May 18-20, 1999. The purpose of the workshop is to present and exchange research results from theoretical and experimental work in combustion science using the reduced-gravity environment as a research tool. The results are contributed by researchers funded by NASA throughout the United States at universities, industry and government research agencies, and by researchers from at least eight international partner countries that are also participating in the microgravity combustion science research discipline. These research results are intended for use by public and private sector organizations for academic purposes, for the development of technologies needed for the Human Exploration and Development of Space, and to improve Earth-bound combustion and fire-safety related technologies.

  7. Sixth International Microgravity Combustion Workshop

    NASA Technical Reports Server (NTRS)

    Sacksteder, Kurt (Compiler)

    2001-01-01

    This conference proceedings document is a compilation of papers presented orally or as poster displays to the Sixth International Microgravity Combustion Workshop held in Cleveland, Ohio on May 22-24, 2001. The purpose of the workshop is to present and exchange research results from theoretical and experimental work in combustion science using the reduced-gravity environment as a research tool. The results are contributed by researchers funded by NASA throughout the United States at universities, industry and government research agencies, and by researchers from international partner countries that are also participating in the microgravity combustion science research discipline. These research results are intended for use by public and private sector organizations for academic purposes, for the development of technologies needed for Human Exploration and Development of Space, and to improve Earth-bound combustion and fire-safety related technologies.

  8. Astrobiology Workshop: Leadership in Astrobiology

    NASA Technical Reports Server (NTRS)

    DeVincenzi, D. (Editor); Briggs, G.; Cohen, M.; Cuzzi, J.; DesMarais, D.; Harper, L.; Morrison, D.; Pohorille, A.

    1996-01-01

    Astrobiology is defined in the 1996 NASA Strategic Plan as 'The study of the living universe.' At NASA's Ames Research Center, this endeavor encompasses the use of space to understand life's origin, evolution, and destiny in the universe. Life's origin refers to understanding the origin of life in the context of the origin and diversity of planetary systems. Life's evolution refers to understanding how living systems have adapted to Earth's changing environment, to the all-pervasive force of gravity, and how they may adapt to environments beyond Earth. Life's destiny refers to making long-term human presence in space a reality, and laying the foundation for understanding and managing changes in Earth's environment. The first Astrobiology Workshop brought together a diverse group of researchers to discuss the following general questions: Where and how are other habitable worlds formed? How does life originate? How have the Earth and its biosphere influenced each other over time? Can terrestrial life be sustained beyond our planet? How can we expand the human presence to Mars? The objectives of the Workshop included: discussing the scope of astrobiology, strengthening existing efforts for the study of life in the universe, identifying new cross-disciplinary programs with the greatest potential for scientific return, and suggesting steps needed to bring this program to reality. Ames has been assigned the lead role for astrobiology by NASA in recognition of its strong history of leadership in multidisciplinary research in the space, Earth, and life sciences and its pioneering work in studies of the living universe. This initial science workshop was established to lay the foundation for what is to become a national effort in astrobiology, with anticipated participation by the university community, other NASA centers, and other agencies. This workshop (the first meeting of its kind ever held) involved life, Earth, and space scientists in a truly interdisciplinary sharing

  9. Merkel Cell Polyomavirus Small T Antigen Mediates Microtubule Destabilization To Promote Cell Motility and Migration

    PubMed Central

    Knight, Laura M.; Stakaityte, Gabriele; Wood, Jennifer, J.; Abdul-Sada, Hussein; Griffiths, David A.; Howell, Gareth J.; Wheat, Rachel; Blair, G. Eric; Steven, Neil M.; Macdonald, Andrew; Blackbourn, David J.

    2014-01-01

    ABSTRACT Merkel cell carcinoma (MCC) is an aggressive skin cancer of neuroendocrine origin with a high propensity for recurrence and metastasis. Merkel cell polyomavirus (MCPyV) causes the majority of MCC cases due to the expression of the MCPyV small and large tumor antigens (ST and LT, respectively). Although a number of molecular mechanisms have been attributed to MCPyV tumor antigen-mediated cellular transformation or replication, to date, no studies have investigated any potential link between MCPyV T antigen expression and the highly metastatic nature of MCC. Here we use a quantitative proteomic approach to show that MCPyV ST promotes differential expression of cellular proteins implicated in microtubule-associated cytoskeletal organization and dynamics. Intriguingly, we demonstrate that MCPyV ST expression promotes microtubule destabilization, leading to a motile and migratory phenotype. We further highlight the essential role of the microtubule-associated protein stathmin in MCPyV ST-mediated microtubule destabilization and cell motility and implicate the cellular phosphatase catalytic subunit protein phosphatase 4C (PP4C) in the regulation of this process. These findings suggest a possible molecular mechanism for the highly metastatic phenotype associated with MCC. IMPORTANCE Merkel cell polyomavirus (MCPyV) causes the majority of cases of Merkel cell carcinoma (MCC), an aggressive skin cancer with a high metastatic potential. However, the molecular mechanisms leading to virally induced cancer development have yet to be fully elucidated. In particular, no studies have investigated any potential link between the virus and the highly metastatic nature of MCC. We demonstrate that the MCPyV small tumor antigen (ST) promotes the destabilization of the host cell microtubule network, which leads to a more motile and migratory cell phenotype. We further show that MCPyV ST induces this process by regulating the phosphorylation status of the cellular microtubule

  10. 1981 NRC/BNL/IEEE standards workshop on human factors and nuclear safety. The man-machine interface and human reliability: an assessment and projection

    SciTech Connect

    Hall, R.E.; Fragola, J.R.; Luckas, W.J. Jr.

    1981-09-01

    The role of the human in the safety of nuclear power plant operations was addressed in a meeting held in Myrtle Beach, SC in August 1981. Presentation were made on Control Room reviews, safety parameter display systems, the integration of human factors in the entire design process, and the use of automated control features. A need was shown for the development of a taxonomy or model to structure future data gathering and the need for models and data to address the issue of cognitive behavior. The primary effect of this behavior on risk was identified. Discussion sessions on the human impact on reliability, and control room design and evaluation were included. (ACR)

  11. Growth Hormone Research Society Workshop Summary: Consensus Guidelines for Recombinant Human Growth Hormone Therapy in Prader-Willi Syndrome

    PubMed Central

    Tony, Michèle; Höybye, Charlotte; Allen, David B.; Tauber, Maïthé; Christiansen, Jens Sandahl; Ambler, Geoffrey R.; Battista, Renaldo; Beauloye, Véronique; Berall, Glenn; Biller, Beverly M. K.; Butler, Merlin G.; Cassidy, Suzanne B.; Chihara, Kazuo; Cohen, Pinchas; Craig, Maria; Farholt, Stense; Goetghebeur, Mireille; Goldstone, Anthony P.; Greggi, Tiziana; Grugni, Graziano; Hokken-Koelega, Anita C.; Johannsson, Gudmundur; Johnson, Keegan; Kemper, Alex; Kopchick, John J.; Malozowski, Saul; Miller, Jennifer; Mogul, Harriette R.; Muscatelli, Françoise; Nergårdh, Ricard; Nicholls, Robert D.; Radovick, Sally; Rosenthal, M. Sara; Sipilä, Ilkka; Tarride, Jean-Eric; Vogels, Annick; Waters, Michael J.

    2013-01-01

    Context: Recombinant human GH (rhGH) therapy in Prader-Willi syndrome (PWS) has been used by the medical community and advocated by parental support groups since its approval in the United States in 2000 and in Europe in 2001. Its use in PWS represents a unique therapeutic challenge that includes treating individuals with cognitive disability, varied therapeutic goals that are not focused exclusively on increased height, and concerns about potential life-threatening adverse events. Objective: The aim of the study was to formulate recommendations for the use of rhGH in children and adult patients with PWS. Evidence: We performed a systematic review of the clinical evidence in the pediatric population, including randomized controlled trials, comparative observational studies, and long-term studies (>3.5 y). Adult studies included randomized controlled trials of rhGH treatment for ≥ 6 months and uncontrolled trials. Safety data were obtained from case reports, clinical trials, and pharmaceutical registries. Methodology: Forty-three international experts and stakeholders followed clinical practice guideline development recommendations outlined by the AGREE Collaboration (www.agreetrust.org). Evidence was synthesized and graded using a comprehensive multicriteria methodology (EVIDEM) (http://bit.ly.PWGHIN). Conclusions: Following a multidisciplinary evaluation, preferably by experts, rhGH treatment should be considered for patients with genetically confirmed PWS in conjunction with dietary, environmental, and lifestyle interventions. Cognitive impairment should not be a barrier to treatment, and informed consent/assent should include benefit/risk information. Exclusion criteria should include severe obesity, uncontrolled diabetes mellitus, untreated severe obstructive sleep apnea, active cancer, or psychosis. Clinical outcome priorities should vary depending upon age and the presence of physical, mental, and social disability, and treatment should be continued for as

  12. Epidemiology of Environmental Exposures and Human Autoimmune Diseases: Findings from a National Institute of Environmental Health Sciences Expert Panel Workshop

    PubMed Central

    Alfredsson, Lars; Costenbader, Karen H.; Kamen, Diane L.; Nelson, Lorene; Norris, Jill M.; De Roos, Anneclaire J.

    2012-01-01

    Autoimmune diseases (AID) are a collection of many complex disorders of unknown etiology resulting in immune responses to self-antigens and are thought to result from interactions between genetic and environmental factors. Here we review the epidemiologic evidence for the role of environmental factors in the development of human AID, the conclusions that can be drawn from the existing data, critical knowledge gaps, and research needed to fill these gaps and to resolve uncertainties. We specifically summarize the state of knowledge and our levels of confidence in the role of specific agents in the development of autoimmune diseases, and we define the areas of greatest impact for future investigations. Among our consensus findings we are confident that: 1) crystalline silica exposure can contribute to the development of several AID; 2) solvent exposure can contribute to the development of systemic sclerosis; 3) smoking can contribute to the development of seropositive rheumatoid arthritis; and 4) an inverse association exists between ultraviolet radiation exposure and the risk of development of multiple sclerosis. We suggest that more studies of phenotypes, genotypes, and multiple exposures are needed. Additional knowledge gaps needing investigation include: defining important windows in the timing of exposures and latencies relating to age, developmental state, and hormonal changes; understanding dose-response relationships; and elucidating mechanisms for disease development. Addressing these essential issues will require more resources to support research, particularly of rare AID, but knowledge of the risks conferred by environmental factors in specific genetic contexts could pave the way for prevention of AID in the future. PMID:22739348

  13. Epidemiology of environmental exposures and human autoimmune diseases: findings from a National Institute of Environmental Health Sciences Expert Panel Workshop.

    PubMed

    Miller, Frederick W; Alfredsson, Lars; Costenbader, Karen H; Kamen, Diane L; Nelson, Lorene M; Norris, Jill M; De Roos, Anneclaire J

    2012-12-01

    Autoimmune diseases (AID) are a collection of many complex disorders of unknown etiology resulting in immune responses to self-antigens and are thought to result from interactions between genetic and environmental factors. Here we review the epidemiologic evidence for the role of environmental factors in the development of human AID, the conclusions that can be drawn from the existing data, critical knowledge gaps, and research needed to fill these gaps and to resolve uncertainties. We specifically summarize the state of knowledge and our levels of confidence in the role of specific agents in the development of autoimmune diseases, and we define the areas of greatest impact for future investigations. Among our consensus findings we are confident that: 1) crystalline silica exposure can contribute to the development of several AID; 2) solvent exposure can contribute to the development of systemic sclerosis; 3) smoking can contribute to the development of seropositive rheumatoid arthritis; and 4) an inverse association exists between ultraviolet radiation exposure and the risk of development of multiple sclerosis. We suggest that more studies of phenotypes, genotypes, and multiple exposures are needed. Additional knowledge gaps needing investigation include: defining important windows in the timing of exposures and latencies relating to age, developmental state, and hormonal changes; understanding dose-response relationships; and elucidating mechanisms for disease development. Addressing these essential issues will require more resources to support research, particularly of rare AID, but knowledge of the risks conferred by environmental factors in specific genetic contexts could pave the way for prevention of AID in the future.

  14. Productivity Workshop Plan.

    ERIC Educational Resources Information Center

    Drewes, Donald W.

    This document presents a plan for conducting productivity workshops sponsored by state vocational education agencies for state agency staff, field vocational educators, and representatives of businesses and industry. The rationale is discussed, and workshop goals and objectives are stated. Suggested procedures are described for the attainment of…

  15. Workshop in Translating Literature

    ERIC Educational Resources Information Center

    Corson, Michael; And Others

    1975-01-01

    A workshop dealing with literature in translation took place in 1974 at the German Department of the University of Cincinnati. This is a report on its procedures and methods. The workshop dealt with discussion of texts, translation of texts, critique of existing translations and interpretation of content. (TL)

  16. Warehouse Sanitation Workshop Handbook.

    ERIC Educational Resources Information Center

    Food and Drug Administration (DHHS/PHS), Washington, DC.

    This workshop handbook contains information and reference materials on proper food warehouse sanitation. The materials have been used at Food and Drug Administration (FDA) food warehouse sanitation workshops, and are selected by the FDA for use by food warehouse operators and for training warehouse sanitation employees. The handbook is divided…

  17. Sensors Workshop summary report

    NASA Technical Reports Server (NTRS)

    1977-01-01

    A review of the efforts of three workshops is presented. The presentation describes those technological developments that would contribute most to sensor subsystem optimization and improvement of NASA's data acquisition capabilities, and summarizes the recommendations of the sensor technology panels from the most recent workshops.

  18. Mental Models in Human-Computer Interaction. Research Issues about What the User of Software Knows. Workshop on Software Human Factors: Users' Mental Models (Washington, District of Columbia, May 15-16, 1984).

    ERIC Educational Resources Information Center

    Carroll, John M., Ed.; Olson, Judith Reitman, Ed.

    This report incorporates a literature review, a workshop paper, and discussion by workshop participants on the current status of research on what the users of computer systems know, and how these different forms of knowledge fit together in learning and performance. It is noted that such research is important to the problem of designing systems…

  19. Ethnic differences in polyomavirus simian virus 40 seroprevalence among women in Houston, Texas

    PubMed Central

    Wong, Connie; Vilchez, Regis A.; Quiroz, Jorge; Adam, Ervin; Butel, Janet S.

    2012-01-01

    Objective To examine the prevalence and distribution among racial/ethnic groups of polyomavirus SV40 antibodies in women in Houston, Texas. Methods Women in three different cohorts reflecting the evolving demographics of Houston were evaluated for frequency of SV40 antibodies using a plaque-reduction neutralization assay. Results Women in cohort A (enrolled 1972–1973) were 68% (145/212) African-American and 32% Caucasian; the overall frequency of SV40 neutralizing antibodies was 7%. Women in cohort B (enrolled 1975–1977) were Caucasian with an overall frequency of SV40 neutralizing antibodies of 18% (37/211). Women in cohort C (enrolled 1993–1995) were 50% (199/400) African-American, 25% Caucasian, and 25% Hispanic; the overall frequency of SV40 neutralizing antibodies was 10%. Logistic regression analysis for cohort A showed no difference in SV40 neutralizing antibodies with respect to race/ethnicity, pregnancy status, number of previous pregnancies, or history of sexually transmitted diseases. For cohort C, race/ethnicity was identified as a significant factor associated with SV40 neutralizing antibodies, with Hispanics having a seroprevalence of 23% compared to 5–6% in the other two groups (p = 0.01). Conclusions A significantly higher SV40 seroprevalence was found among Hispanics than other racial/ethnic groups in the city of Houston. Findings are compatible with a model that certain population groups potentially exposed to SV40-contaminated oral poliovaccines have maintained cycles of SV40 infections. PMID:22940252

  20. Emerging From the Unknown: Structural and Functional Features of Agnoprotein of Polyomaviruses

    PubMed Central

    Saribas, A. Sami; Coric, Pascale; Hamazaspyan, Anahit; Davis, William; Axman, Rachel; White, Martyn K.; Abou-Gharbia, Magid; Childers, Wayne; Condra, Jon H.; Bouaziz, Serge; Safak, Mahmut

    2016-01-01

    Agnoprotein is an important regulatory protein of polyomaviruses, including JCV, BKV, and SV40. In the absence of its expression, these viruses are unable to sustain their productive life cycle. It is a highly basic phosphoprotein that localizes mostly to the perinuclear area of infected cells, although a small amount of the protein is also found in nucleus. Much has been learned about the structure and function of this important regulatory protein in recent years. It forms highly stable dimers/oligomers in vitro and in vivo through its Leu/Ile/Phe-rich domain. Structural NMR studies revealed that this domain adopts an alpha-helix conformation and plays a critical role in the stability of the protein. It associates with cellular proteins, including YB-1, p53, Ku70, FEZ1, HP1α, PP2A, AP-3, PCNA, and α-SNAP; and viral proteins, including small t antigen, large T antigen, HIV-1 Tat, and JCV VP1; and significantly contributes the viral transcription and replication. This review summarizes the recent advances in the structural and functional properties of this important regulatory protein. PMID:26831433

  1. A longitudinal study on avian polyomavirus-specific antibodies in captive Spix's macaws (Cyanopsitta spixii).

    PubMed

    Deb, Amrita; Foldenauer, Ulrike; Borjal, Raffy Jim; Streich, W Jürgen; Lüken, Caroline; Johne, Reimar; Müller, Hermann; Hammer, Sven

    2010-09-01

    Avian polyomavirus (APV) causes a range of disease syndromes in psittacine birds, from acute fatal disease to subclinical infections, depending on age, species, and other unidentified risk factors. To determine the prevalence of APV-specific antibodies in a captive population of Spix's macaws (Cyanopsitta spixii) in Quatar, 54 birds were tested by blocking enzyme-linked immunosorbent assay. A prevalence of 48.1% for APV antibodies, which indicates viral exposure, was found. Of 36 Spix's macaws that were serially tested over a period of 4 years, 50.0% were consistently positive, 36.1% were consistently negative, 5.5% had permanently declining antibody levels, and 2.8% showed variable results. By using polymerase chain reaction testing on whole blood samples, an apparent viremia was detected in 1 of 44 birds (2.3%), although contamination provides a likely explanation for this isolated positive result in a hand-reared chick. The white blood cell count was significantly higher in antibody-positive birds compared with antibody-negative birds (P < .05). Because antibody-positive and antibody-negative birds were housed together without a change in their respective antibody status, transmission of APV within the adult breeding population appeared to be a rare event.

  2. Whole genome sequence of a goose haemorrhagic polyomavirus detected in Hungary.

    PubMed

    Fehér, Enikő; Lengyel, György; Dán, Adám; Farkas, Szilvia L; Bányai, Krisztián

    2014-06-01

    Goose haemorrhagic polyomavirus (GHPV) provoke haemorrhagic nephritis and enteritis of domestic geese. Outbreaks were detected in European countries and caused economic losses for goose keepers. Domestic ducks may be infected with GHPV without any signs typical for geese. The genomic organisation of some isolates was described but the gene functions and the pathomechanisms of the virus was not precisely defined. Here we describe the genome sequence and structure of GHPV of a goose from a Hungarian goose flock showing characteristics of the haemorrhagic nephritis and enteritis. The GHPV genome investigated in this study was 5252 bp long and was very similar (99% nucleotide identity) to sequences deposited in the GenBank. All the whole GHPV genomes possess the same ORFs in length, including the VP1, VP2, VP3, ORF-X, t and T tumour antigens. Amino acid changes are detected mainly in the putative ORF-X region. Data about the GHPV genome imply a conserved genomic structure among isolates from different countries. Genomic and epidemiological studies may help vaccine development efforts and identify potential heterologous reservoirs of GHPV.

  3. Murine Polyomavirus Cell Surface Receptors Activate Distinct Signaling Pathways Required for Infection

    PubMed Central

    O’Hara, Samantha D.

    2016-01-01

    ABSTRACT Virus binding to the cell surface triggers an array of host responses, including activation of specific signaling pathways that facilitate steps in virus entry. Using mouse polyomavirus (MuPyV), we identified host signaling pathways activated upon virus binding to mouse embryonic fibroblasts (MEFs). Pathways activated by MuPyV included the phosphatidylinositol 3-kinase (PI3K), FAK/SRC, and mitogen-activated protein kinase (MAPK) pathways. Gangliosides and α4-integrin are required receptors for MuPyV infection. MuPyV binding to both gangliosides and the α4-integrin receptors was required for activation of the PI3K pathway; however, either receptor interaction alone was sufficient for activation of the MAPK pathway. Using small-molecule inhibitors, we confirmed that the PI3K and FAK/SRC pathways were required for MuPyV infection, while the MAPK pathway was dispensable. Mechanistically, the PI3K pathway was required for MuPyV endocytosis, while the FAK/SRC pathway enabled trafficking of MuPyV along microtubules. Thus, MuPyV interactions with specific cell surface receptors facilitate activation of signaling pathways required for virus entry and trafficking. Understanding how different viruses manipulate cell signaling pathways through interactions with host receptors could lead to the identification of new therapeutic targets for viral infection. PMID:27803182

  4. Merkel Cell Polyomavirus Large T Antigen Disrupts Host Genomic Integrity and Inhibits Cellular Proliferation

    PubMed Central

    Li, Jing; Wang, Xin; Diaz, Jason; Tsang, Sabrina H.; Buck, Christopher B.

    2013-01-01

    Clonal integration of Merkel cell polyomavirus (MCV) DNA into the host genome has been observed in at least 80% of Merkel cell carcinoma (MCC). The integrated viral genome typically carries mutations that truncate the C-terminal DNA binding and helicase domains of the MCV large T antigen (LT), suggesting a selective pressure to remove this MCV LT region during tumor development. In this study, we show that MCV infection leads to the activation of host DNA damage responses (DDR). This activity was mapped to the C-terminal helicase-containing region of the MCV LT. The MCV LT-activated DNA damage kinases, in turn, led to enhanced p53 phosphorylation, upregulation of p53 downstream target genes, and cell cycle arrest. Compared to the N-terminal MCV LT fragment that is usually preserved in mutants isolated from MCC tumors, full-length MCV LT shows a decreased potential to support cellular proliferation, focus formation, and anchorage-independent cell growth. These apparently antitumorigenic effects can be reversed by a dominant-negative p53 inhibitor. Our results demonstrate that MCV LT-induced DDR activates p53 pathway, leading to the inhibition of cellular proliferation. This study reveals a key difference between MCV LT and simian vacuolating virus 40 LT, which activates a DDR but inhibits p53 function. This study also explains, in part, why truncation mutations that remove the MCV LT C-terminal region are necessary for the oncogenic progression of MCV-associated cancers. PMID:23760247

  5. Antibodies to merkel cell polyomavirus T antigen oncoproteins reflect tumor burden in merkel cell carcinoma patients.

    PubMed

    Paulson, Kelly G; Carter, Joseph J; Johnson, Lisa G; Cahill, Kevin W; Iyer, Jayasri G; Schrama, David; Becker, Juergen C; Madeleine, Margaret M; Nghiem, Paul; Galloway, Denise A

    2010-11-01

    Merkel cell polyomavirus (MCPyV) is a common infectious agent that is likely involved in the etiology of most Merkel cell carcinomas (MCC). Serum antibodies recognizing the MCPyV capsid protein VP1 are detectable at high titer in nearly all MCC patients and remain stable over time. Although antibodies to the viral capsid indicate prior MCPyV infection, they provide limited clinical insight into MCC because they are also detected in more than half of the general population. We investigated whether antibodies recognizing MCPyV large and small tumor-associated antigens (T-Ag) would be more specifically associated with MCC. Among 530 population control subjects, these antibodies were present in only 0.9% and were of low titer. In contrast, among 205 MCC cases, 40.5% had serum IgG antibodies that recognize a portion of T-Ag shared between small and large T-Ags. Among cases, titers of T-Ag antibodies fell rapidly (∼8-fold per year) in patients whose cancer did not recur, whereas they rose rapidly in those with progressive disease. Importantly, in several patients who developed metastases, the rise in T-Ag titer preceded clinical detection of disease spread. These results suggest that antibodies recognizing T-Ag are relatively specifically associated with MCC, do not effectively protect against disease progression, and may serve as a clinically useful indicator of disease status.

  6. Pathological and epidemiological significance of goose haemorrhagic polyomavirus infection in ducks.

    PubMed

    Corrand, Léni; Gelfi, Jacqueline; Albaric, Olivier; Etievant, Mélanie; Pingret, Jean-Luc; Guerin, Jean-Luc

    2011-08-01

    Goose haemorrhagic polyomavirus (GHPV) is the viral agent of haemorrhagic nephritis enteritis of geese, a lethal disease of goslings. It was recently shown that GHPV can also be detected in Muscovy and mule ducks. The goal of the present study was to investigate the pathobiology of GHPV in ducks. In the first experiment, field isolates of GHPV from Muscovy or mule ducks were fully sequenced and compared with goose GHPV. These duck isolates were then used to inoculate 1-day-old goslings. Typical clinical signs and lesions of haemorrhagic nephritis enteritis of geese were reproduced, indicating that "duck-GHPV" isolates are virulent in geese. In the second experiment, 1-day-old and 21-day-old Muscovy ducklings were infected by a reference GHPV strain. In both cases, neither clinical signs nor histopathological lesions were observed. However, the virus was detected in cloacal bursae and sera, and serological responses were detected at 12 days post infection. These findings suggest firstly that one common genotype of GHPV circulates among ducks and geese, and secondly that ducks may be infected by GHPV but show no pathologic evidence of infection, whereas geese express clinical signs. GHPV infection should therefore be considered as being carried in ducks and of epidemiological relevance in cases of contact with goose flocks.

  7. T-antigen-independent replication of polyomavirus DNA in murine embryonal carcinoma cells

    SciTech Connect

    Dandolo, L.; Aghion, J.; Blangy, D.

    1984-02-01

    Expression of wild-type polyomavirus (Py) is restricted in murine embryonal carcinoma (EC) cells. The block appears to be located at the level of early transcription. Since no T antigen is produced, the authors investigated the fate of viral DNA upon infection of these cells; they showed that wild-type Py DNA replicates efficiently in all EC cells, probably via a T-antigen-independent mechanism. Furthermore, they studied, at permissive and restrictive temperatures, the replication of tsa (thermosensitive for T antigen) viral DNA of an in vitro-constructed deletion mutant lacking part of the early region coding sequences and of a double mutant carrying both the tsa mutation and the PyEC F9 mutation (allowing expression of early and late viral functions in EC cells). The results imply that replication of wild-type A2 strain Py DNA can occur in EC cells in the absence of a functional T antigen. However, this protein clearly enhances viral DNA replication and is absolutely required in differentiated cells.

  8. Detection of KI WU and Merkel cell polyomavirus in respiratory tract of cystic fibrosis patients.

    PubMed

    Iaria, M; Caccuri, F; Apostoli, P; Giagulli, C; Pelucchi, F; Padoan, R F; Caruso, A; Fiorentini, S

    2015-06-01

    In the last few years, many reports have confirmed the presence of WU, KI and Merkel cell (MC) polyomaviruses (PyV) in respiratory samples wordwide, but their pathogenic role in patients with underlying conditions such as cystic fibrosis is still debated. To determine the prevalence of MCPyV, WUPyV and KIPyV, we conducted a 1-year-long microbiological testing of respiratory specimens from 93 patients with cystic fibrosis in Brescia, Italy. We detected PyV DNA in 94 out of 337 analysed specimens. KIPyV was the most common virus detected (12.1%), followed by WUPyV (8.9%) and MCPyV (6.8%). We found an intriguing association between the presence of MCPyV and the concurrent isolation of Pseudomonas aeruginosa, as well as with the patient status, classified as chronically colonized with P. aeruginosa. Our study adds perspective on the prevalence and the potential pathogenic role of PyV infections.

  9. Identification of a polyomavirus microRNA highly expressed in tumors.

    PubMed

    Chen, Chun Jung; Cox, Jennifer E; Azarm, Kristopher D; Wylie, Karen N; Woolard, Kevin D; Pesavento, Patricia A; Sullivan, Christopher S

    2015-02-01

    Polyomaviruses (PyVs) are associated with tumors including Merkel cell carcinoma (MCC). Several PyVs encode microRNAs (miRNAs) but to date no abundant PyV miRNAs have been reported in tumors. To better understand the function of the Merkel cell PyV (MCPyV) miRNA, we examined phylogenetically-related viruses for miRNA expression. We show that two primate PyVs and the more distantly-related raccoon PyV (RacPyV) encode miRNAs that share genomic position and partial sequence identity with MCPyV miRNAs. Unlike MCPyV miRNA in MCC, RacPyV miRNA is highly abundant in raccoon tumors. RacPyV miRNA negatively regulates reporters of early viral (T antigen) transcripts, yet robust viral miRNA expression is tolerated in tumors. We also identify raccoon miRNAs expressed in RacPyV-associated neuroglial brain tumors, including several likely oncogenic miRNAs (oncomiRs). This work describes the first PyV miRNA abundantly expressed in tumors and is consistent with a possible role for both host and viral miRNAs in RacPyV-associated tumors.

  10. Antibody response to BK polyomavirus as a prognostic biomarker and potential therapeutic target in prostate cancer

    PubMed Central

    Keller, Xavier Etienne; Kardas, Piotr; Acevedo, Claudio; Sais, Giovanni; Poyet, Cédric; Banzola, Irina; Mortezavi, Ashkan; Seifert, Burkhardt; Sulser, Tullio

    2015-01-01

    Infectious agents, including the BK polyomavirus (BKPyV), have been proposed as important inflammatory pathogens in prostate cancer. Here, we evaluated whether the preoperative antibody response to BKPyV large T antigen (LTag) and viral capsid protein 1 (VP1) was associated with the risk of biochemical recurrence in 226 patients undergoing radical prostatectomy for primary prostate cancer. Essentially, the multivariate Cox regression analysis revealed that preoperative seropositivity to BKPyV LTag significantly reduced the risk of biochemical recurrence, independently of established predictors of biochemical recurrence such as tumor stage, Gleason score and surgical margin status. The predictive accuracy of the regression model was denotatively increased by the inclusion of the BKPyV LTag serostatus. In contrast, the VP1 serostatus was of no prognostic value. Finally, the BKPyV LTag serostatus was associated with a peculiar cytokine gene expression profile upon assessment of the cellular immune response elicited by LTag. Taken together, our findings suggest that the BKPyV LTag serology may serve as a prognostic factor in prostate cancer. If validated in additional studies, this biomarker may allow for better treatment decisions after radical prostatectomy. Finally, the favorable outcome of LTag seropositive patients may provide a potential opportunity for novel therapeutic approaches targeting a viral antigen. PMID:25749042

  11. The association between polyomavirus BK strains and BKV viruria in liver transplant recipients

    PubMed Central

    Wang, Robert Y. L.; Li, Yi-Jung; Lee, Wei-Chen; Wu, Hsin-Hsu; Lin, Chan-Yu; Lee, Cheng-Chia; Chen, Yung-Chang; Hung, Cheng-Chieh; Yang, Chih-Wei; Tian, Ya-Chung

    2016-01-01

    BK virus (BKV) is a polyomavirus that cause of allograft dysfunction among kidney transplant recipients. The role of BKV infection in non-renal solid organ transplant recipients is not well understood neither for the relationship between various BKV strains with occurrence of BKV viral viruria. This study aimed to understand the prevalence of BKV infection and identified of BKV various strains in the urine of liver transplant recipients. There was not significant difference of renal outcome between high BKV viruria and low BKV viruria in the liver transplant recipients. The WW-non-coding control region (NCCR) BKV detected in urine was associated with higher urinary BKV load, whereas the Dunlop-NCCR BKV was detected in the urine of low urinary BKV load. An in vitro cultivation system demonstrated that WW-BKV strain exhibiting the higher viral DNA replication efficiency and higher BKV load. Altogether, this is the first study to demonstrate the impact of BKV strains on the occurrence of BK viruria in the liver transplant recipients. PMID:27338010

  12. Conference report: interdisciplinary workshop in the philosophy of medicine: parentalism and trust.

    PubMed

    Bullock, Emma; Gergel, Tania; Kingma, Elselijn

    2015-06-01

    On 13 June 2014, the Centre for the Humanities and Health at King's College London hosted a 1-day workshop on 'parentalism and trust'. This workshop was the sixth in a series of workshops whose aim is to provide a new model for high-quality open interdisciplinary engagement between medical professionals and philosophers. This report briefly describes the workshop methodology and the discussions on the day.

  13. Detection of polyomavirus simian virus 40 tumor antigen DNA in AIDS-related systemic non-Hodgkin lymphoma

    NASA Technical Reports Server (NTRS)

    Vilchez, Regis A.; Lednicky, John A.; Halvorson, Steven J.; White, Zoe S.; Kozinetz, Claudia A.; Butel, Janet S.

    2002-01-01

    Systemic non-Hodgkin lymphoma (S-NHL) is a common malignancy during HIV infection, and it is hypothesized that infectious agents may be involved in the etiology. Epstein-Barr virus DNA is found in <40% of patients with AIDS-related S-NHL, suggesting that other oncogenic viruses, such as polyomaviruses, may play a role in pathogenesis. We analyzed AIDS-related S-NHL samples, NHL samples from HIV-negative patients, peripheral blood leukocytes from HIV-infected and -uninfected patients without NHL, and lymph nodes without tumors from HIV-infected patients. Specimens were examined by polymerase chain reaction analysis with use of primers specific for an N-terminal region of the oncoprotein large tumor antigen ( T-ag ) gene conserved among all three polyomaviruses (simian virus 40 [SV40], JC virus, and BK virus). Polyomavirus T-ag DNA sequences, proven to be SV40-specific, were detected more frequently in AIDS-related S-NHL samples (6 of 26) than in peripheral blood leukocytes from HIV-infected patients (6 of 26 vs. 0 of 69; p =.0001), NHL samples from HIV-negative patients (6 of 26 vs. 0 of 10; p =.09), or lymph nodes (6 of 26 vs. 0 of 7; p =.16). Sequences of C-terminal T-ag DNA from SV40 were amplified from two AIDS-related S-NHL samples. Epstein-Barr virus DNA sequences were detected in 38% (10 of 26) AIDS-related S-NHL samples, 50% (5 of 10) HIV-negative S-NHL samples, and 57% (4 of 7) lymph nodes. None of the S-NHL samples were positive for both Epstein-Barr virus DNA and SV40 DNA. Further studies of the possible role of SV40 in the pathogenesis of S-NHL are warranted.

  14. Ocean margins workshop

    SciTech Connect

    1990-12-31

    The Department of Energy (DOE) is announcing the refocusing of its marine research program to emphasize the study of ocean margins and their role in modulating, controlling, and driving Global Change phenomena. This is a proposal to conduct a workshop that will establish priorities and an implementation plan for a new research initiative by the Department of Energy on the ocean margins. The workshop will be attended by about 70 scientists who specialize in ocean margin research. The workshop will be held in the Norfolk, Virginia area in late June 1990.

  15. Soil Moisture Workshop

    NASA Technical Reports Server (NTRS)

    Heilman, J. L. (Editor); Moore, D. G. (Editor); Schmugge, T. J. (Editor); Friedman, D. B. (Editor)

    1978-01-01

    The Soil Moisture Workshop was held at the United States Department of Agriculture National Agricultural Library in Beltsville, Maryland on January 17-19, 1978. The objectives of the Workshop were to evaluate the state of the art of remote sensing of soil moisture; examine the needs of potential users; and make recommendations concerning the future of soil moisture research and development. To accomplish these objectives, small working groups were organized in advance of the Workshop to prepare position papers. These papers served as the basis for this report.

  16. Nuclear Innovation Workshops Report

    SciTech Connect

    Jackson, John Howard; Allen, Todd Randall; Hildebrandt, Philip Clay; Baker, Suzanne Hobbs

    2015-09-01

    The Nuclear Innovation Workshops were held at six locations across the United States on March 3-5, 2015. The data collected during these workshops has been analyzed and sorted to bring out consistent themes toward enhancing innovation in nuclear energy. These themes include development of a test bed and demonstration platform, improved regulatory processes, improved communications, and increased public-private partnerships. This report contains a discussion of the workshops and resulting themes. Actionable steps are suggested at the end of the report. This revision has a small amount of the data in Appendix C removed in order to avoid potential confusion.

  17. Human polyomavirus JCV late leader peptide region contains important regulatory elements

    SciTech Connect

    Akan, Ilhan; Sariyer, Ilker Kudret; Biffi, Renato; Palermo, Victoria; Woolridge, Stefanie; White, Martyn K.; Amini, Shohreh |; Khalili, Kamel; Safak, Mahmut . E-mail: msafak@temple.edu

    2006-05-25

    Transcription is a complex process that relies on the cooperative interaction between sequence-specific factors and the basal transcription machinery. The strength of a promoter depends on upstream or downstream cis-acting DNA elements, which bind transcription factors. In this study, we investigated whether DNA elements located downstream of the JCV late promoter, encompassing the late leader peptide region, which encodes agnoprotein, play regulatory roles in the JCV lytic cycle. For this purpose, the entire coding region of the leader peptide was deleted and the functional consequences of this deletion were analyzed. We found that viral gene expression and replication were drastically reduced. Gene expression also decreased from a leader peptide point mutant but to a lesser extent. This suggested that the leader peptide region of JCV might contain critical cis-acting DNA elements to which transcription factors bind and regulate viral gene expression and replication. We analyzed the entire coding region of the late leader peptide by a footprinting assay and identified three major regions (region I, II and III) that were protected by nuclear proteins. Further investigation of the first two protected regions by band shift assays revealed a new band that appeared in new infection cycles, suggesting that viral infection induces new factors that interact with the late leader peptide region of JCV. Analysis of the effect of the leader peptide region on the promoter activity of JCV by transfection assays demonstrated that this region has a positive and negative effect on the large T antigen (LT-Ag)-mediated activation of the viral early and late promoters, respectively. Furthermore, a partial deletion analysis of the leader peptide region encompassing the protected regions I and II demonstrated a significant down-regulation of viral gene expression and replication. More importantly, these results were similar to that obtained from a complete deletion of the late leader peptide region, indicating the critical importance of these two protected regions in JCV regulation. Altogether, these findings suggest that the late leader peptide region contains important regulatory elements to which transcription factors bind and contribute to the JCV gene regulation and replication.

  18. Biomedical Polar Research Workshop Minutes

    NASA Technical Reports Server (NTRS)

    1990-01-01

    This workshop was conducted to provide a background of NASA and National Science Foundation goals, an overview of previous and current biomedical research, and a discussion about areas of potential future joint activities. The objectives of the joint research were: (1) to develop an understanding of the physiological, psychological, and behavioral alterations and adaptations to extreme environments of the polar regions; (2) to ensure the health, well-being, and performance of humans in these environments; and (3) to promote the application of biomedical research to improve the quality of life in all environments.

  19. From Workshop to Classroom: Bridging GIS Success

    ERIC Educational Resources Information Center

    Stonier, Francis; Hong, Jung Eun

    2016-01-01

    This article shares the first-time geographic information system (GIS) experiences of two advanced placement human geography classes. The teacher had participated in a summer GIS workshop and then brought those skills into her classroom for the students' benefit. Eighteen students shared their experiences researching their family history, working…

  20. Workshop overview: Arsenic research and risk assessment

    SciTech Connect

    Sams, Reeder Wolf, Douglas C.; Ramasamy, Santhini; Ohanian, Ed; Chen, Jonathan; Lowit, Anna

    2007-08-01

    The chronic exposure of humans through consumption of high levels of inorganic arsenic (iAs)-contaminated drinking water is associated with skin lesions, peripheral vascular disease, hypertension, and cancers. Additionally, humans are exposed to organic arsenicals when used as pesticides and herbicides (e.g., monomethylarsonic acid, dimethylarsinic acid (DMA{sup V}) also known as cacodylic acid). Extensive research has been conducted to characterize the adverse health effects that result from exposure to iAs and its metabolites to describe the biological pathway(s) that lead to adverse health effects. To further this effort, on May 31, 2006, the United States Environmental Protection Agency (USEPA) sponsored a meeting entitled 'Workshop on Arsenic Research and Risk Assessment'. The invited participants from government agencies, academia, independent research organizations and consultants were asked to present their current research. The overall focus of these research efforts has been to determine the potential human health risks due to environmental exposures to arsenicals. Pursuant in these efforts is the elucidation of a mode of action for arsenicals. This paper provides a brief overview of the workshop goals, regulatory context for arsenical research, mode of action (MOA) analysis in human health risk assessment, and the application of MOA analysis for iAs and DMA{sup V}. Subsequent papers within this issue will present the research discussed at the workshop, ensuing discussions, and conclusions of the workshop.

  1. Learning by Sharing in the Outdoors. Workshop of the University of New Hampshire Outdoor Education Program (Durham, New Hampshire, March 5, 1983).

    ERIC Educational Resources Information Center

    Gass, Michael A.

    A collection of information from the "Learning by Sharing in the Outdoors" workshop provides brief summaries of each workshop session along with lists of human, community, and bibliographical resources available to outdoor education practitioners, the workshop itinerary, and promotional brochures. The 18 workshop topics included:…

  2. Urban Waters Workshop

    EPA Pesticide Factsheets

    This page will house information leading up to the 2017 Urban Waters National Training Workshop. The agenda, hotel and other quarterly updates will be posted to this page including information about how to register.

  3. Workshop: Teaching Primitive Arts.

    ERIC Educational Resources Information Center

    Jordison, Jerry

    1999-01-01

    Discusses the concrete and spiritual aspects of teaching workshops on survival skills or primitive arts. Gives details on lostproofing, or ways to teach a child not to get lost in the outdoors; building a survival shelter; and wilderness cooking. (CDS)

  4. The Career Development Workshop

    ERIC Educational Resources Information Center

    Marsh, P. J.

    1973-01-01

    This article describes a career planning workshop for managers and its purpose is to support and accelerate the process of individual development without organizational coercion or manipulation. (Author/RK)

  5. An Aerospace Workshop

    ERIC Educational Resources Information Center

    Hill, Bill

    1972-01-01

    Describes the 16-day, 10,000 mile national tour of the nation's major aerospace research and development centers by 65 students enrolled in Central Washington State College's Summer Aerospace Workshop. (Author/MB)

  6. Colorado Model Rocketry Workshop.

    ERIC Educational Resources Information Center

    Galindez, Peter

    1978-01-01

    Describes a summer workshop course in rocketry offered to educators and sponsored by industry. The participants built various model rockets and equipment and worked on challenging practical problems and activities. (GA)

  7. Special parallel processing workshop

    SciTech Connect

    1994-12-01

    This report contains viewgraphs from the Special Parallel Processing Workshop. These viewgraphs deal with topics such as parallel processing performance, message passing, queue structure, and other basic concept detailing with parallel processing.

  8. Cybernetics and Workshop Design.

    ERIC Educational Resources Information Center

    Eckstein, Daniel G.

    1979-01-01

    Cybernetic sessions allow for the investigation of several variables concurrently, resulting in a large volume of input compacted into a concise time frame. Three session questions are reproduced to illustrate the variety of ideas generated relative to workshop design. (Author)

  9. ISIS Workshops Using Virtualization

    NASA Astrophysics Data System (ADS)

    Becker, K. J.; Becker, T. L.

    2015-06-01

    ISIS workshops are now using virtualization technology to improve the user experience and create a stable, consistent and useful ISIS installation for educational purposes as well as future processing needs.

  10. Transportation Management Workshop: Proceedings

    SciTech Connect

    Not Available

    1993-10-01

    This report is a compilation of discussions presented at the Transportation Management Workshop held in Gaithersburg, Maryland. Topics include waste packaging, personnel training, robotics, transportation routing, certification, containers, and waste classification.

  11. Complex Flow Workshop Report

    SciTech Connect

    none,

    2012-05-01

    This report documents findings from a workshop on the impacts of complex wind flows in and out of wind turbine environments, the research needs, and the challenges of meteorological and engineering modeling at regional, wind plant, and wind turbine scales.

  12. Appalachian Stream Mitigation Workshop

    EPA Pesticide Factsheets

    A 5 day workshop in 2011 developed for state and federal regulatory and resource agencies, who review, comment on and/or approve compensatory mitigation plans for surface coal mining projects in Appalachia

  13. INTEGRATED RISK ASSESSMENT - RESULTS FROM AN INTERNATIONAL WORKSHOP

    EPA Science Inventory

    The WHO International Programme on Chemical Safety and international partners have developed a framework for integrated assessment of human health and ecological risks and four case studies. An international workshop was convened to consider how ecological and health risk assess...

  14. TCGA Workshop: Genomics and Biology of Glioblastoma Multiforme (GBM) - TCGA

    Cancer.gov

    The National Cancer Institute (NCI) and National Human Genome Research Institute (NHGRI) held a workshop entitled, “Genomics and Biology of Glioblastoma Multiforme (GBM),” to review the initial GBM data from the TCGA pilot project.

  15. Workshop I: Gender Studies

    NASA Astrophysics Data System (ADS)

    Hennessey, Eden; Kurup, Anitha; Meza-Montes, Lilia; Shastri, Prajval; Ghose, Shohini

    2015-12-01

    Participants in the Gender Studies workshop of the 5th IUPAP International Conference on Women in Physics discussed the gender question in science practice from a policy perspective, informed by investigations from the social science disciplines. The workshop's three sessions—"Equity and Education: Examining Gender Stigma in Science," "A Comparative Study of Women Scientists and Engineers: Experiences in India and the US," and "Toward Gender Equity Through Policy: Characterizing the Social Impact of Interventions—are summarized, and the resulting recommendations presented.

  16. Space Mechanisms Technology Workshop

    NASA Technical Reports Server (NTRS)

    Oswald, Fred B. (Editor)

    2001-01-01

    The Mechanical Components Branch at NASA Glenn Research Center hosted a workshop to discuss the state of drive systems technology needed for space exploration. The Workshop was held Thursday, November 2, 2000. About 70 space mechanisms experts shared their experiences from working in this field and considered technology development that will be needed to support future space exploration in the next 10 to 30 years.

  17. OEXP Analysis Tools Workshop

    NASA Technical Reports Server (NTRS)

    Garrett, L. Bernard; Wright, Robert L.; Badi, Deborah; Findlay, John T.

    1988-01-01

    This publication summarizes the software needs and available analysis tools presented at the OEXP Analysis Tools Workshop held at the NASA Langley Research Center, Hampton, Virginia on June 21 to 22, 1988. The objective of the workshop was to identify available spacecraft system (and subsystem) analysis and engineering design tools, and mission planning and analysis software that could be used for various NASA Office of Exploration (code Z) studies, specifically lunar and Mars missions.

  18. Space Mechanisms Technology Workshop

    NASA Technical Reports Server (NTRS)

    Oswald, Fred B. (Editor)

    2002-01-01

    The Mechanical Components Branch at NASA Glenn Research Center hosted a workshop on Tuesday, May 14, 2002, to discuss space mechanisms technology. The theme for this workshop was 'Working in the Cold,' a focus on space mechanisms that must operate at low temperatures. We define 'cold' as below -60C (210 K), such as would be found near the equator of Mars. However, we are also concerned with much colder temperatures such as in permanently dark craters of the Moon (about 40 K).

  19. Industrial Fuel Flexibility Workshop

    SciTech Connect

    none,

    2006-09-01

    On September 28, 2006, in Washington, DC, ITP and Booz Allen Hamilton conducted a fuel flexibility workshop with attendance from various stakeholder groups. Workshop participants included representatives from the petrochemical, refining, food and beverage, steel and metals, pulp and paper, cement and glass manufacturing industries; as well as representatives from industrial boiler manufacturers, technology providers, energy and waste service providers, the federal government and national laboratories, and developers and financiers.

  20. Visual detection of goose haemorrhagic polyomavirus in geese and ducks by loop-mediated isothermal amplification.

    PubMed

    Woźniakowski, Grzegorz; Tarasiuk, Karolina

    2015-01-01

    Goose haemorrhagic polyomavirus (GHPV) is an aetiological agent of haemorrhagic nephritis and enteritis of geese occurring in geese (Anser anser). GHPV may also infect Muscovy ducks (Carina mochata) and mule ducks. Early detection of GHPV is important to isolate the infected birds from the rest of the flock thus limiting infection transmission. The current diagnosis of haemorrhagic nephritis and enteritis of geese is based on virus isolation, histopathological examination, haemagglutination inhibition assay, ELISA and polymerase chain reaction (PCR). Recently, real-time PCR assay was developed which considerably improved detection of GHPV. In spite of many advantages, these methods are still time-consuming and inaccessible for laboratories with limited access to ELISA plate readers or PCR thermocyclers. The aim of our study was to develop loop-mediated isothermal amplification (LAMP) that may be conducted in a water bath. Two pairs of specific primers complementary to VP1 gene of GHPV were designed. The results of GHPV LAMP were recorded under ultraviolet light. Our study showed LAMP was able to specifically amplify VP1 fragment of a GHPV without cross-reactivity with other pathogens of geese and ducks. LAMP detected as little as 1.5 pg of DNA extracted from a GHPV standard strain (150 pg/µl). The optimized LAMP was used to examine 18 field specimens collected from dead and clinically diseased geese and ducks aged from 1 to 12 weeks. The positive signal for GHPV was detected in three out of 18 (16.6%) specimens. These results were reproducible and consistent with those of four real-time PCR. To the best of our knowledge this is the first report on LAMP application for the GHPV detection.

  1. Banff Initiative for Quality Assurance in Transplantation (BIFQUIT): Reproducibility of Polyomavirus Immunohistochemistry in Kidney Allografts

    PubMed Central

    Adam, Benjamin; Randhawa, Parmjeet; Chan, Samantha; Zeng, Gang; Regele, Heinz; Kushner, Yael B.; Colvin, Robert B.; Reeve, Jeff; Mengel, Michael

    2014-01-01

    Immunohistochemistry is the gold standard for diagnosing (positive versus negative) polyomavirus BK (BKV) nephropathy and has the potential for disease staging based on staining intensity and quantification of infected cells. This multicenter trial evaluated the reproducibility of BKV immunohistochemistry among 81 pathologists at 60 institutions. Participants stained tissue microarray slides and scored them for staining intensity and percentage of positive nuclei. Staining protocol details and evaluation scores were collected online. Slides were returned for centralized panel re-evaluation and kappa statistics were calculated. Individual assessment of staining intensity and percentage was more reproducible than combined scoring. Inter-institutional reproducibility was moderate for staining intensity (κ=0.49) and percentage (κ=0.42), fair for combined (κ=0.25), and best for simple positive/negative scoring (κ=0.63). Inter-observer reproducibility was substantial for intensity (κ=0.74), percentage (κ=0.66), and positive/negative (κ=0.67), and moderate for combined scoring (κ=0.43). Inter-laboratory reproducibility was fair for intensity (κ=0.37), percentage (κ=0.40), and combined (κ=0.24), but substantial for positive/negative scoring (κ=0.78). BKV RNA copies/cell correlated with staining intensity (r=0.56) and percentage (r=0.62). These results indicate that BKV immunohistochemistry is reproducible between observers but scoring should be simplified to a single-feature schema. Standardization of tissue processing and staining protocols would further improve inter-laboratory reproducibility. PMID:25091177

  2. Polyomavirus JCV excretion and genotype analysis in HIV-infected patients receiving highly active antiretroviral therapy

    NASA Technical Reports Server (NTRS)

    Lednicky, John A.; Vilchez, Regis A.; Keitel, Wendy A.; Visnegarwala, Fehmida; White, Zoe S.; Kozinetz, Claudia A.; Lewis, Dorothy E.; Butel, Janet S.

    2003-01-01

    OBJECTIVE: To assess the frequency of shedding of polyomavirus JC virus (JCV) genotypes in urine of HIV-infected patients receiving highly active antiretroviral therapy (HAART). METHODS: Single samples of urine and blood were collected prospectively from 70 adult HIV-infected patients and 68 uninfected volunteers. Inclusion criteria for HIV-infected patients included an HIV RNA viral load < 1000 copies, CD4 cell count of 200-700 x 106 cells/l, and stable HAART regimen. PCR assays and sequence analysis were carried out using JCV-specific primers against different regions of the virus genome. RESULTS: JCV excretion in urine was more common in HIV-positive patients but not significantly different from that of the HIV-negative group [22/70 (31%) versus 13/68 (19%); P = 0.09]. HIV-positive patients lost the age-related pattern of JCV shedding (P = 0.13) displayed by uninfected subjects (P = 0.01). Among HIV-infected patients significant differences in JCV shedding were related to CD4 cell counts (P = 0.03). Sequence analysis of the JCV regulatory region from both HIV-infected patients and uninfected volunteers revealed all to be JCV archetypal strains. JCV genotypes 1 (36%) and 4 (36%) were the most common among HIV-infected patients, whereas type 2 (77%) was the most frequently detected among HIV-uninfected volunteers. CONCLUSION: These results suggest that JCV shedding is enhanced by modest depressions in immune function during HIV infection. JCV shedding occurred in younger HIV-positive persons than in the healthy controls. As the common types of JCV excreted varied among ethnic groups, JCV genotypes associated with progressive multifocal leukoencephalopathy may reflect demographics of those infected patient populations.

  3. Clinical and prognostic significance of Merkel cell polyomavirus in nonsmall cell lung cancer

    PubMed Central

    Kim, Gun-Jik; Lee, Jae-Ho; Lee, Deok Heon

    2017-01-01

    Abstract Recently, an association between Merkel cell polyomavirus (MCPyV) and epidermal growth factor receptor (EGFR) mutations in nonsmall cell lung cancer (NSCLC) was reported. However, the underlying carcinogenic effects and the prognosis related to MCPyV are still unclear. The aim of this study was to clarify the incidence and prognosis related to MCPyV infections in NSCLC. Tissue samples from 167 NSCLC patients (92 with squamous cell carcinomas [SCCs] and 75 with adenocarcinomas) were analyzed for the presence of MCPyV and EGFR mutations. Clinicopathological characteristics, disease-free survival rate, and overall survival rate were assessed with respect to MCPyV. MCPyV DNA was detected in 30 patients (18.0%) out of 167 patients, and EGFR mutations were found in 31 out of 127 patients (24.4%). EGFR mutations were more frequently detected in MCPyV-positive patients than in MCPyV-negative patients; however, this did not reach statistical significance (P = 0.075). There was no difference in overall survival between patients with and without MCPyV infections. The disease-free survival rate of patients with pN0 stage, SCC, or EGFR mutations was lower for patients with MCPyV than without MCPyV (P = 0.036, 0.042, and 0.050, respectively). Although the prevalence of MCPyV infection was relatively low, the presence of MCPyV DNA was significantly correlated with cancer prognosis in subgroups of NSCLC patients. These results suggest that MCPyV may be partly associated with pathogenesis and prognosis in some cases of NSCLC. PMID:28099328

  4. Blue native protein electrophoresis for studies of mouse polyomavirus morphogenesis and interactions between the major capsid protein VP1 and cellular proteins.

    PubMed

    Horníková, Lenka; Man, Petr; Forstová, Jitka

    2011-12-01

    Morphogenesis of the mouse polyomavirus virion is a complex and not yet well understood process. Nuclear lysates of infected cells and cells transiently producing the major capsid protein (VP1) of the mouse polyomavirus and whole-cell lysates were separated by blue native polyacrylamide gel electrophoresis (BN-PAGE) to characterize the participation of cellular proteins in virion precursor complexes. Several VP1-specific complexes were found by immunostaining with the anti-VP1 antibody. Some of these complexes contained proteins from the heat shock protein 70 family. The BN-PAGE was found to be a useful tool for the identification of protein complexes by immunostaining of separated cell lysates. However, whole-cell lysates and lysates of isolated nuclei of cells infected with polyomavirus appeared to be too complex for BN-PAGE separation followed by mass spectrometry. No distinct bands specific for cells infected with polyomavirus were detected by Coomassie blue stained gels, hence this method is not suitable for the discovery of new cellular proteins participating in virion assembly. Nevertheless, BN-PAGE can be valuable for the analyses of different types of complexes formed by proteins after their enrichment or isolation by affinity chromatography.

  5. JC Polyomavirus Abundance and Distribution in Progressive Multifocal Leukoencephalopathy (PML) Brain Tissue Implicates Myelin Sheath in Intracerebral Dissemination of Infection

    PubMed Central

    Wharton, Keith A.; Quigley, Catherine; Themeles, Marian; Dunstan, Robert W.; Doyle, Kathryn; Cahir-McFarland, Ellen; Wei, Jing; Buko, Alex; Reid, Carl E.; Sun, Chao; Carmillo, Paul; Sur, Gargi; Carulli, John P.; Mansfield, Keith G.; Westmoreland, Susan V.; Staugaitis, Susan M.; Fox, Robert J.; Meier, Werner; Goelz, Susan E.

    2016-01-01

    Over half of adults are seropositive for JC polyomavirus (JCV), but rare individuals develop progressive multifocal leukoencephalopathy (PML), a demyelinating JCV infection of the central nervous system. Previously, PML was primarily seen in immunosuppressed patients with AIDS or certain cancers, but it has recently emerged as a drug safety issue through its association with diverse immunomodulatory therapies. To better understand the relationship between the JCV life cycle and PML pathology, we studied autopsy brain tissue from a 70-year-old psoriasis patient on the integrin alpha-L inhibitor efalizumab following a ~2 month clinical course of PML. Sequence analysis of lesional brain tissue identified PML-associated viral mutations in regulatory (non-coding control region) DNA, capsid protein VP1, and the regulatory agnoprotein, as well as 9 novel mutations in capsid protein VP2, indicating rampant viral evolution. Nine samples, including three gross PML lesions and normal-appearing adjacent tissues, were characterized by histopathology and subject to quantitative genomic, proteomic, and molecular localization analyses. We observed a striking correlation between the spatial extent of demyelination, axonal destruction, and dispersion of JCV along white matter myelin sheath. Our observations in this case, as well as in a case of PML-like disease in an immunocompromised rhesus macaque, suggest that long-range spread of polyomavirus and axonal destruction in PML might involve extracellular association between virus and the white matter myelin sheath. PMID:27191595

  6. Survey of molecular chaperone requirement for the biosynthesis of hamster polyomavirus VP1 protein in Saccharomyces cerevisiae.

    PubMed

    Valaviciute, Monika; Norkiene, Milda; Goda, Karolis; Slibinskas, Rimantas; Gedvilaite, Alma

    2016-07-01

    A number of viruses utilize molecular chaperones during various stages of their life cycle. It has been shown that members of the heat-shock protein 70 (Hsp70) chaperone family assist polyomavirus capsids during infection. However, the molecular chaperones that assist the formation of recombinant capsid viral protein 1 (VP1)-derived virus-like particles (VLPs) in yeast remain unclear. A panel of yeast strains with single chaperone gene deletions were used to evaluate the chaperones required for biosynthesis of recombinant hamster polyomavirus capsid protein VP1. The impact of deletion or mild overexpression of chaperone genes was determined in live cells by flow cytometry using enhanced green fluorescent protein (EGFP) fused with VP1. Targeted genetic analysis demonstrated that VP1-EGFP fusion protein levels were significantly higher in yeast strains in which the SSZ1 or ZUO1 genes encoding ribosome-associated complex components were deleted. The results confirmed the participation of cytosolic Hsp70 chaperones and suggested the potential involvement of the Ydj1 and Caj1 co-chaperones and the endoplasmic reticulum chaperones in the biosynthesis of VP1 VLPs in yeast. Likewise, the markedly reduced levels of VP1-EGFP in Δhsc82 and Δhsp82 yeast strains indicated that both Hsp70 and Hsp90 chaperones might assist VP1 VLPs during protein biosynthesis.

  7. Ultrastructural and molecular confirmation of the trichodysplasia spinulosa-associated polyomavirus in biopsies of patients with trichodysplasia spinulosa.

    PubMed

    Elaba, Zendee; Hughey, Lauren; Isayeva, Tatyana; Weeks, Beth; Solovan, Caius; Solovastru, Laura; Andea, Aleodor

    2012-11-01

    Trichodysplasia spinulosa (TS) is a rare and only recently characterized cutaneous disease occurring in immunocompromised patients. The disease is characterized by spiny follicular papules on clinical examination and by the presence of viral inclusions at ultrastructural examination. In the last year, this virus has been identified as a new member of the polyomavirus family and designated as TS-associated polyomavirus (TSPyV). We report two organ transplant patients with this disease in which we were able to identify the TSPyV at ultrastructural and molecular level from formalin-fixed paraffin-embedded biopsies of lesional skin. Similar to prior described cases, the patients presented with follicular papules which were concentrated on the central face and associated with alopecia. Histopathology of both cases showed dilated follicular infundibula plugged with cornified eosinophilic cells containing large trichohyaline granules. Transmission electron microscopy on paraffin-embedded tissue in case 1 showed 28-nm intracellular viral particles morphologically consistent with polyoma virus. For both cases the presence of TSPyV was confirmed by polymerase chain reaction with virus-specific primers followed by identification by direct sequencing. These two cases show the presence of the newly described TSPyV in TS further establishing its association with this distinctive disease.

  8. Investigation of host-cell influence on polyomavirus biological activity and the major virus-coded structural protein VP1

    SciTech Connect

    Ludlow, J.W.

    1987-01-01

    The host cell influence on polyomavirus biological activity and modification of the major virus-coded structural protein VP1 was investigated. Polyoma virions resulting from the permissive infections of primary mouse kidney cells, as compared to virus progeny from primary mouse embryo cells, exhibited a ten-fold greater ability to agglutinate guinea pig erythrocytes, a three-fold lower ability to become internalized into monopincytotic vesicles, and a two-fold lower ability to initiate a productive infection based on positive nuclear immunofluorescence when using mouse embryo host cells. Mouse kidney cell progeny were found to bind to host cells less specifically than mouse-embryo cell progeny. Two-dimensional gel electrophoresis of VP1, following in vivo labeling with /sup 32/P, revealed that species D and E of the mouse kidney cell progeny were phosphorylated to the same degree while mouse embryo cell progeny VP1 species E and F were phosphorylated equally. In vivo labeling of polyomavirus with /sup 35/S-sulfate, followed by gel electrophoretic analysis of the structural proteins and two-dimensional chromatographic analysis of the VP1 amino acids revealed that species E and F are modified by sulfation of tyrosine. These data suggest that host cells play a role in modulating biological activity of the virus by affecting the degree and site specific modification of the major capsid protein VP1. Modification of this protein may influence the recognition of virus attachment proteins for specific cellular receptors as well as other biological functions.

  9. Prevalence of Papillomaviruses, Polyomaviruses, and Herpesviruses in Triple-Negative and Inflammatory Breast Tumors from Algeria Compared with Other Types of Breast Cancer Tumors

    PubMed Central

    Corbex, Marilys; Bouzbid, Sabiha; Traverse-Glehen, Alexandra; Aouras, Hayette; McKay-Chopin, Sandrine; Carreira, Christine; Lankar, Abdelaziz; Tommasino, Massimo; Gheit, Tarik

    2014-01-01

    Background The possible role of viruses in breast cancer etiology remains an unresolved question. We hypothesized that if some viruses are involved, it may be in a subgroup of breast cancers only. Epidemiological arguments drove our interest in breast cancer subgroups that are more frequent in Africa, namely inflammatory breast cancer (IBC) and triple-negative breast cancer. We tested whether viral prevalence was significantly higher in these subgroups. Materials and Methods One hundred fifty-five paraffin-embedded malignant breast tumors were randomly selected at the pathology laboratory of the University Hospital of Annaba (Algeria) to include one third of IBC and two thirds of non-IBC. They were tested for the presence of DNA from 61 viral agents (46 human papillomaviruses, 10 polyomaviruses, and 5 herpesviruses) using type-specific multiplex genotyping assays, which combine multiplex PCR and bead-based Luminex technology. Results Viral DNA was found in 22 (17.9%) of 123 tumors. The most prevalent viruses were EBV1 and HPV16. IBC tumors carried significantly more viruses (any type) than non-IBC tumors (30% vs. 13%, p<0.04). Similarly, triple-negative tumors displayed higher virus-positivity than non-triple-negative tumors (44% vs. 14%, p<0.009). Conclusions Our results suggest an association between the presence of viral DNA and aggressive breast cancer phenotypes (IBC, triple-negative). While preliminary, they underline the importance of focusing on subgroups when studying viral etiology in breast cancer. Further studies on viruses in breast cancer should be conducted in much larger samples to confirm these initial findings. PMID:25478862

  10. BK Polyomavirus Replication in Renal Tubular Epithelial Cells Is Inhibited by Sirolimus, but Activated by Tacrolimus Through a Pathway Involving FKBP-12.

    PubMed

    Hirsch, H H; Yakhontova, K; Lu, M; Manzetti, J

    2016-03-01

    BK polyomavirus (BKPyV) replication causes nephropathy and premature kidney transplant failure. Insufficient BKPyV-specific T cell control is regarded as a key mechanism, but direct effects of immunosuppressive drugs on BKPyV replication might play an additional role. We compared the effects of mammalian target of rapamycin (mTOR)- and calcineurin-inhibitors on BKPyV replication in primary human renal tubular epithelial cells. Sirolimus impaired BKPyV replication with a 90% inhibitory concentration of 4 ng/mL by interfering with mTOR-SP6-kinase activation. Sirolimus inhibition was rapid and effective up to 24 h postinfection during viral early gene expression, but not thereafter, during viral late gene expression. The mTORC-1 kinase inhibitor torin-1 showed a similar inhibition profile, supporting the notion that early steps of BKPyV replication depend on mTOR activity. Cyclosporine A also inhibited BKPyV replication, while tacrolimus activated BKPyV replication and reversed sirolimus inhibition. FK binding protein 12kda (FKBP-12) siRNA knockdown abrogated sirolimus inhibition and increased BKPyV replication similar to adding tacrolimus. Thus, sirolimus and tacrolimus exert opposite effects on BKPyV replication in renal tubular epithelial cells by a mechanism involving FKBP-12 as common target. Immunosuppressive drugs may therefore contribute directly to the risk of BKPyV replication and nephropathy besides suppressing T cell functions. The data provide rationales for clinical trials aiming at reducing the risk of BKPyV replication and disease in kidney transplantation.

  11. BK Polyomavirus Replication in Renal Tubular Epithelial Cells Is Inhibited by Sirolimus, but Activated by Tacrolimus Through a Pathway Involving FKBP‐12

    PubMed Central

    Yakhontova, K.; Lu, M.; Manzetti, J.

    2015-01-01

    BK polyomavirus (BKPyV) replication causes nephropathy and premature kidney transplant failure. Insufficient BKPyV‐specific T cell control is regarded as a key mechanism, but direct effects of immunosuppressive drugs on BKPyV replication might play an additional role. We compared the effects of mammalian target of rapamycin (mTOR)‐ and calcineurin‐inhibitors on BKPyV replication in primary human renal tubular epithelial cells. Sirolimus impaired BKPyV replication with a 90% inhibitory concentration of 4 ng/mL by interfering with mTOR–SP6‐kinase activation. Sirolimus inhibition was rapid and effective up to 24 h postinfection during viral early gene expression, but not thereafter, during viral late gene expression. The mTORC‐1 kinase inhibitor torin‐1 showed a similar inhibition profile, supporting the notion that early steps of BKPyV replication depend on mTOR activity. Cyclosporine A also inhibited BKPyV replication, while tacrolimus activated BKPyV replication and reversed sirolimus inhibition. FK binding protein 12kda (FKBP‐12) siRNA knockdown abrogated sirolimus inhibition and increased BKPyV replication similar to adding tacrolimus. Thus, sirolimus and tacrolimus exert opposite effects on BKPyV replication in renal tubular epithelial cells by a mechanism involving FKBP‐12 as common target. Immunosuppressive drugs may therefore contribute directly to the risk of BKPyV replication and nephropathy besides suppressing T cell functions. The data provide rationales for clinical trials aiming at reducing the risk of BKPyV replication and disease in kidney transplantation. PMID:26639422

  12. International Lighting in Controlled Environments Workshop

    NASA Technical Reports Server (NTRS)

    Tibbits, Ted W. (Editor)

    1994-01-01

    Lighting is a central and critical aspect of control in environmental research for plant research and is gaining recognition as a significant factor to control carefully for animal and human research. Thus this workshop was convened to reevaluate the technology that is available today and to work toward developing guidelines for the most effective use of lighting in controlled environments with emphasis on lighting for plants but also to initiate interest in the development of improved guidelines for human and animal research.

  13. IFPA meeting 2014 workshop report: Animal models to study pregnancy pathologies; new approaches to study human placental exposure to xenobiotics; biomarkers of pregnancy pathologies; placental genetics and epigenetics; the placenta and stillbirth and fetal growth restriction.

    PubMed

    Barbaux, S; Erwich, J J H M; Favaron, P O; Gil, S; Gallot, D; Golos, T G; Gonzalez-Bulnes, A; Guibourdenche, J; Heazell, A E P; Jansson, T; Laprévote, O; Lewis, R M; Miller, R K; Monk, D; Novakovic, B; Oudejans, C; Parast, M; Peugnet, P; Pfarrer, C; Pinar, H; Roberts, C T; Robinson, W; Saffery, R; Salomon, C; Sexton, A; Staff, A C; Suter, M; Tarrade, A; Wallace, J; Vaillancourt, C; Vaiman, D; Worton, S A; Lash, G E

    2015-04-01

    Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialized topics. At IFPA meeting 2014 there were six themed workshops, five of which are summarized in this report. These workshops related to various aspects of placental biology but collectively covered areas of animal models, xenobiotics, pathological biomarkers, genetics and epigenetics, and stillbirth and fetal growth restriction.

  14. The QED Workshop

    SciTech Connect

    Pieper, G.W.

    1994-07-01

    On May 18--20, 1994, Argonne National Laboratory hosted the QED Workshop. The workshop was supported by special funding from the Office of Naval Research. The purpose of the workshop was to assemble of a group of researchers to consider whether it is desirable and feasible to build a proof-checked encyclopedia of mathematics, with an associated facility for theorem proving and proof checking. Among the projects represented were Coq, Eves, HOL, ILF, Imps, MathPert, Mizar, NQTHM, NuPrl, OTTER, Proof Pad, Qu-Prolog, and RRL. Although the content of the QED project is highly technical rigorously proof-checked mathematics of all sorts the discussions at the workshop were rarely technical. No prepared talks or papers were given. Instead, the discussions focused primarily on such political, sociological, practical, and aesthetic questions, such as Why do it? Who are the customers? How can one get mathematicians interested? What sort of interfaces are desirable? The most important conclusion of the workshop was that QED is an idea worthy pursuing, a statement with which virtually all the participants agreed. In this document, the authors capture some of the discussions and outline suggestions for the start of a QED scientific community.

  15. t4 Workshop Report*

    PubMed Central

    Kleensang, Andre; Maertens, Alexandra; Rosenberg, Michael; Fitzpatrick, Suzanne; Lamb, Justin; Auerbach, Scott; Brennan, Richard; Crofton, Kevin M.; Gordon, Ben; Fornace, Albert J.; Gaido, Kevin; Gerhold, David; Haw, Robin; Henney, Adriano; Ma’ayan, Avi; McBride, Mary; Monti, Stefano; Ochs, Michael F.; Pandey, Akhilesh; Sharan, Roded; Stierum, Rob; Tugendreich, Stuart; Willett, Catherine; Wittwehr, Clemens; Xia, Jianguo; Patton, Geoffrey W.; Arvidson, Kirk; Bouhifd, Mounir; Hogberg, Helena T.; Luechtefeld, Thomas; Smirnova, Lena; Zhao, Liang; Adeleye, Yeyejide; Kanehisa, Minoru; Carmichael, Paul; Andersen, Melvin E.; Hartung, Thomas

    2014-01-01

    Summary Despite wide-spread consensus on the need to transform toxicology and risk assessment in order to keep pace with technological and computational changes that have revolutionized the life sciences, there remains much work to be done to achieve the vision of toxicology based on a mechanistic foundation. A workshop was organized to explore one key aspect of this transformation – the development of Pathways of Toxicity (PoT) as a key tool for hazard identification based on systems biology. Several issues were discussed in depth in the workshop: The first was the challenge of formally defining the concept of a PoT as distinct from, but complementary to, other toxicological pathway concepts such as mode of action (MoA). The workshop came up with a preliminary definition of PoT as “A molecular definition of cellular processes shown to mediate adverse outcomes of toxicants”. It is further recognized that normal physiological pathways exist that maintain homeostasis and these, sufficiently perturbed, can become PoT. Second, the workshop sought to define the adequate public and commercial resources for PoT information, including data, visualization, analyses, tools, and use-cases, as well as the kinds of efforts that will be necessary to enable the creation of such a resource. Third, the workshop explored ways in which systems biology approaches could inform pathway annotation, and which resources are needed and available that can provide relevant PoT information to the diverse user communities. PMID:24127042

  16. 77 FR 31371 - Public Workshop: Privacy Compliance Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-25

    ... compliance fundamentals, privacy and data security, and the privacy compliance life cycle. A learning lunch... SECURITY Office of the Secretary Public Workshop: Privacy Compliance Workshop AGENCY: Privacy Office, DHS. ACTION: Notice Announcing Public Workshop. SUMMARY: The Department of Homeland Security Privacy...

  17. Conducting Effective Staff Development Workshops

    ERIC Educational Resources Information Center

    Bishop, Kay; Janczak, Sue

    2005-01-01

    Staff development workshops conducted by library media specialists can assist teachers to integrate information literacy skills and technology into their curricula. Guidelines are presented on the planning and implementation of such workshops.

  18. Measurement control workshop instructional materials

    SciTech Connect

    Gibbs, Philip; Crawford, Cary; McGinnis, Brent

    2014-04-01

    A workshop to teach the essential elements of an effective nuclear materials control and accountability (MC&A) programs are outlined, along with the modes of Instruction, and the roles and responsibilities of participants in the workshop.

  19. Thin film solar cell workshop

    NASA Technical Reports Server (NTRS)

    Armstrong, Joe; Jeffrey, Frank

    1993-01-01

    A summation of responses to questions posed to the thin-film solar cell workshop and the ensuing discussion is provided. Participants in the workshop included photovoltaic manufacturers (both thin film and crystalline), cell performance investigators, and consumers.

  20. Space Station Workstation Technology Workshop Report

    NASA Technical Reports Server (NTRS)

    Moe, K. L.; Emerson, C. M.; Eike, D. R.; Malone, T. B.

    1985-01-01

    This report describes the results of a workshop conducted at Goddard Space Flight Center (GSFC) to identify current and anticipated trends in human-computer interface technology that may influence the design or operation of a space station workstation. The workshop was attended by approximately 40 persons from government and academia who were selected for their expertise in some aspect of human-machine interaction research. The focus of the workshop was a 1 1/2 brainstorming/forecasting session in which the attendees were assigned to interdisciplinary working groups and instructed to develop predictions for each of the following technology areas: (1) user interface, (2) resource management, (3) control language, (4) data base systems, (5) automatic software development, (6) communications, (7) training, and (8) simulation. This report is significant in that it provides a unique perspective on workstation design for the space station. This perspective, which is characterized by a major emphasis on user requirements, should be most valuable to Phase B contractors involved in design development of the space station workstation. One of the more compelling results of the workshop is the recognition that no major technological breakthroughs are required to implement the current workstation concept. What is required is the creative application of existing knowledge and technology.

  1. Report on the consensus workshop on formaldehyde

    SciTech Connect

    Gough, M.; Hart, R.; Karrh, B.W.; Koestner, A.; Neal, R.; Parkinson, D.; Perera, F.; Powell, K.E.; Rosenkranz, S.

    1984-01-01

    The Consensus Workshop on Formaldehyde consisted of bringing together scientists from academia, government, industry and public interest groups to address some important toxicological questions concerning the health effects of formaldehyde. The participants in the workshop, the Executive Panel which coordinated the meeting, and the questions posed, all were chosen through a broadly based nomination process in order to achieve as comprehensive a consensus as possible. The subcommittees considered the toxicological problems associated with formaldehyde in the areas of exposure, epidemiology, carcinogenicity/histology/genotoxicity, immunology/sensitization/irritation, structure activity/biochemistry metabolism, reproduction/teratology, behavior/neurotoxicity/psychology and risk estimation. Some questions considered included the possible human carcinogenicity of formaldehyde, as well as other human health effects, and the interpretation of pathology induced by formaldehyde. These reports, plus introductory material on the procedures used in setting up the Consensus Workshop are presented here. Additionally, there is included a listing of the data base that was made available to the panel chairmen prior to the meeting and was readily accessible to the participants during their deliberations in the meeting. This data base, since it was computerized, was also capable of being searched for important terms. These materials were supplemented by information brought by the panelists. The workshop has defined the consensus concerning a number of major points in formaldehyde toxicology and has identified a number of major deficits in understanding which are important guides to future research. 264 references.

  2. Report on the Consensus Workshop on Formaldehyde.

    PubMed Central

    1984-01-01

    The Consensus Workshop on Formaldehyde consisted of bringing together scientists from academia, government, industry and public interest groups to address some important toxicological questions concerning the health effects of formaldehyde. The participants in the workshop, the Executive Panel which coordinated the meeting, and the questions posed, all were chosen through a broadly based nomination process in order to achieve as comprehensive a consensus as possible. The subcommittees considered the toxicological problems associated with formaldehyde in the areas of exposure, epidemiology, carcinogenicity/histology/genotoxicity, immunology/sensitization/irritation, structure activity/biochemistry/metabolism, reproduction/teratology, behavior/neurotoxicity/psychology and risk estimation. Some questions considered included the possible human carcinogenicity of formaldehyde, as well as other human health effects, and the interpretation of pathology induced by formaldehyde. These reports, plus introductory material on the procedures used in setting up the Consensus Workshop are presented here. Additionally, there is included a listing of the data base that was made available to the panel chairmen prior to the meeting and was readily accessible to the participants during their deliberations in the meeting. This data base, since it was computerized, was also capable of being searched for important terms. These materials were supplemented by information brought by the panelists. The workshop has defined the consensus concerning a number of major points in formaldehyde toxicology and has identified a number of major deficits in understanding which are important guides to future research. PMID:6525992

  3. An International Workshop on Primary Science. Report on the Primary Science Workshop Held after the Conference in Science and Technology Education and Future Human Needs (Bangalore, India, August 1985).

    ERIC Educational Resources Information Center

    Harlen, Wynne, Comp.

    A conference on science and technology and future human needs was attended by over 300 science educators from 64 countries. Educators with particular interest in primary science and technology education extended their stay for an additional seminar. This report highlights the events of that seminar. Contents include: (1) recent and on-going work…

  4. Skylab Orbiter Workshop Illustration

    NASA Technical Reports Server (NTRS)

    1972-01-01

    This cutaway illustration shows the characteristics and basic elements of the Skylab Orbiter Workshop (OWS). The OWS was divided into two major compartments. The lower level provided crew accommodations for sleeping, food preparation and consumption, hygiene, waste processing and disposal, and performance of certain experiments. The upper level consisted of a large work area and housed water storage tanks, a food freezer, storage vaults for film, scientific airlocks, mobility and stability experiment equipment, and other experimental equipment. The compartment below the crew quarters was a container for liquid and solid waste and trash accumulated throughout the mission. A solar array, consisting of two wings covered on one side with solar cells, was mounted outside the workshop to generate electrical power to augment the power generated by another solar array mounted on the solar observatory. Thrusters were provided at one end of the workshop for short-term control of the attitude of the space station.

  5. Workshop on molecular animation.

    PubMed

    Bromberg, Sarina; Chiu, Wah; Ferrin, Thomas E

    2010-10-13

    From February 25 to 26, 2010, in San Francisco, the Resource for Biocomputing, Visualization, and Informatics (RBVI) and the National Center for Macromolecular Imaging (NCMI) hosted a molecular animation workshop for 21 structural biologists, molecular animators, and creators of molecular visualization software. Molecular animation aims to visualize scientific understanding of biomolecular processes and structures. The primary goal of the workshop was to identify the necessary tools for producing high-quality molecular animations, understanding complex molecular and cellular structures, creating publication supplementary materials and conference presentations, and teaching science to students and the public. Another use of molecular animation emerged in the workshop: helping to focus scientific inquiry about the motions of molecules and enhancing informal communication within and between laboratories.

  6. Final Scientific EFNUDAT Workshop

    ScienceCinema

    None

    2016-07-12

    The Final Scientific EFNUDAT Workshop - organized by the CERN/EN-STI group on behalf of n_TOF Collaboration - will be held at CERN, Geneva (Switzerland) from 30 August to 2 September 2010 inclusive.EFNUDAT website: http://www.efnudat.euTopics of interest include: Data evaluationCross section measurementsExperimental techniquesUncertainties and covariancesFission propertiesCurrent and future facilities  International Advisory Committee: C. Barreau (CENBG, France)T. Belgya (IKI KFKI, Hungary)E. Gonzalez (CIEMAT, Spain)F. Gunsing (CEA, France)F.-J. Hambsch (IRMM, Belgium)A. Junghans (FZD, Germany)R. Nolte (PTB, Germany)S. Pomp (TSL UU, Sweden) Workshop Organizing Committee: Enrico Chiaveri (Chairman)Marco CalvianiSamuel AndriamonjeEric BerthoumieuxCarlos GuerreroRoberto LositoVasilis Vlachoudis Workshop Assistant: Géraldine Jean

  7. Workshop on Molecular Animation

    PubMed Central

    Bromberg, Sarina; Chiu, Wah; Ferrin, Thomas E.

    2011-01-01

    Summary February 25–26, 2010, in San Francisco, the Resource for Biocomputing, Visualization and Informatics (RBVI) and the National Center for Macromolecular Imaging (NCMI) hosted a molecular animation workshop for 21 structural biologists, molecular animators, and creators of molecular visualization software. Molecular animation aims to visualize scientific understanding of biomolecular processes and structures. The primary goal of the workshop was to identify the necessary tools for: producing high quality molecular animations, understanding complex molecular and cellular structures, creating publication supplementary materials and conference presentations, and teaching science to students and the public. Another use of molecular animation emerged in the workshop: helping to focus scientific inquiry about the motions of molecules and enhancing informal communication within and between laboratories. PMID:20947014

  8. In-House EVM Workshop

    DTIC Science & Technology

    2007-11-02

    Government Accounting • Management Information System • Workshop Recommendations EVM In- House Workshop Findings • EVM implementation within DoD...Management Command Mr. William “Bill” Gibson Mr. Dominic A. “Chip” Thomas IN- HOUSE ( GOVERNMENT ) EVMS WORKSHOP VALIDATION & SURVEILLANCE D:\\PPT...2Defense Contract Management Command IN- HOUSE ( GOVERNMENT ) EVMS WORKSHOP VALIDATION • WHY VALIDATE/CERTIFY • WHO PAYS THE COST • FACILITY

  9. Cognition in Space Workshop. 1; Metrics and Models

    NASA Technical Reports Server (NTRS)

    Woolford, Barbara; Fielder, Edna

    2005-01-01

    "Cognition in Space Workshop I: Metrics and Models" was the first in a series of workshops sponsored by NASA to develop an integrated research and development plan supporting human cognition in space exploration. The workshop was held in Chandler, Arizona, October 25-27, 2004. The participants represented academia, government agencies, and medical centers. This workshop addressed the following goal of the NASA Human System Integration Program for Exploration: to develop a program to manage risks due to human performance and human error, specifically ones tied to cognition. Risks range from catastrophic error to degradation of efficiency and failure to accomplish mission goals. Cognition itself includes memory, decision making, initiation of motor responses, sensation, and perception. Four subgoals were also defined at the workshop as follows: (1) NASA needs to develop a human-centered design process that incorporates standards for human cognition, human performance, and assessment of human interfaces; (2) NASA needs to identify and assess factors that increase risks associated with cognition; (3) NASA needs to predict risks associated with cognition; and (4) NASA needs to mitigate risk, both prior to actual missions and in real time. This report develops the material relating to these four subgoals.

  10. The Astronomy Workshop

    NASA Astrophysics Data System (ADS)

    Hamilton, D. P.; Asbury, M. L.; Proctor, A.

    2001-12-01

    The Astronomy Workshop (http://janus.astro.umd.edu) is an interactive online astronomy resource developed, and maintained at the University of Maryland, for use by students, educators and the general public. The Astronomy Workshop has been extensively tested and used successfully at many different levels, including High School and Junior High School science classes, University introductory astronomy courses, and University intermediate and advanced astronomy courses. Some topics currently covered in the Astronomy Workshop are: Animated Orbits of Planets and Moons: The orbits of the nine planets and 91 known planetary satellites are shown in animated, to-scale drawings. The orbiting bodies move at their correct relative speeds about their parent, which is rendered as an attractive, to-scale gif image. Solar System Collisions: This most popular of our applications shows what happens when an asteroid or comet with user-defined size and speed impacts a given planet. The program calculates many effects, including the country impacted (if Earth is the target), energy of the explosion, crater size, magnitude of the planetquake generated. It also displays a relevant image (e.g. terrestrial crater, lunar crater, etc.). Planetary and Satellite Data Calculators: These tools allow the user to easily calculate physical data for all of the planets or satellites simultaneously, making comparison very easy. Orbital Simulations: These tools allow the student to investigate different aspects of the three-body problem of celestial mechanics. Astronomy Workshop Bulletin Board: Get innovative teaching ideas and read about in-class experiences with the Astronomy Workshop. Share your ideas with other educators by posting on the Bulletin Board. Funding for the Astronomy Workshop is provided by the National Science Foundation.

  11. The Astronomy Workshop

    NASA Astrophysics Data System (ADS)

    Hamilton, D. P.; Asbury, M. L.

    2000-05-01

    The Astronomy Workshop (http://janus.astro.umd.edu) is an interactive online astronomy resource developed and maintained at the University of Maryland for use by students, educators and the general public. The Astronomy Workshop has been extensively tested and used successfully at many different levels, including High School and Junior High School science classes, University introductory astronomy courses, and University intermediate and advanced astronomy courses. Some topics currently covered in the Astronomy Workshop are: ANIMATED ORBITS OF PLANETS AND MOONS: The orbits of the nine planets and 63 known planetary satellites are shown in animated, to-scale drawings. The orbiting bodies move at their correct relative speeds about their parent, which is rendered as an attractive, to-scale gif image. SOLAR SYSTEM COLLISIONS: This most popular of our applications shows what happens when an asteroid or comet with user-defined size and speed impacts a given planet. The program calculates many effects, including the country impacted (if Earth is the target), energy of explosion, crater size, and magnitude of the ``planetquake'' generated. It also displays a relevant image (e.g. terrestrial crater, lunar crater, etc.). SCALE OF THE UNIVERSE: Travel away from the Earth at a chosen speed and see how long it takes to reach other planets, stars and galaxies. This tool helps students visualize astronomical distances in an intuitive way. SCIENTIFIC NOTATION: Students are interactively guided through conversions between scientific notation and regular numbers. ORBITAL SIMULATIONS: These tools allow the student to investigate different aspects of the three-body problem of celestial mechanics. ASTRONOMY WORKSHOP BULLETIN BOARD: Get innovative teaching ideas and read about in-class experiences with the Astronomy Workshop. Share your ideas with other educators by posting on the Bulletin Board. Funding for the Astronomy Workshop is provided by NSF.

  12. The Astronomy Workshop

    NASA Astrophysics Data System (ADS)

    Hamilton, D. P.; Asbury, M. L.

    1999-12-01

    The Astronomy Workshop (http://janus.astro.umd.edu) is an interactive online astronomy resource developed and maintained at the University of Maryland for use by students, educators and the general public. The Astronomy Workshop has been extensively tested and used successfully at many different levels, including High School and Junior High School science classes, University introductory astronomy courses, and University intermediate and advanced astronomy courses. Some topics currently covered in the Astronomy Workshop are: Animated Orbits of Planets and Moons: The orbits of the nine planets and 63 known planetary satellites are shown in animated, to-scale drawings. The orbiting bodies move at their correct relative speeds about their parent, which is rendered as an attractive, to-scale gif image. Solar System Collisions: This most popular of our applications shows what happens when an asteroid or comet with user-defined size and speed impacts a given planet. The program calculates many effects, including the country impacted (if Earth is the target), energy of explosion, crater size, and magnitude of the ``planetquake'' generated. It also displays a relevant image (e.g. terrestrial crater, lunar crater, etc.). Scale of the Universe: Travel away from the Earth at a chosen speed and see how long it takes to reach other planets, stars and galaxies. This tool helps students visualize astronomical distances in an intuitive way. Scientific Notation: Students are interactively guided through conversions between scientific notation and regular numbers. Orbital Simulations: These tools allow the student to investigate different aspects of the three-body problem of celestial mechanics. Astronomy Workshop Bulletin Board: Get innovative teaching ideas and read about in-class experiences with the Astronomy Workshop. Share your ideas with other educators by posting on the Bulletin Board. Funding for the Astronomy Workshop is provided by NSF.

  13. The Astronomy Workshop

    NASA Astrophysics Data System (ADS)

    Hamilton, D. P.; Asbury, M. L.

    1999-09-01

    The Astronomy Workshop (http://janus.astro.umd.edu) is an interactive online astronomy resource developed and maintained at the University of Maryland for use by students, educators and the general public. The Astronomy Workshop has been extensively tested and used successfully at many different levels, including High School and Junior High School science classes, University introductory astronomy courses, and University intermediate and advanced astronomy courses. Some topics currently covered in the Astronomy Workshop are: Animated Orbits of Planets and Moons: The orbits of the nine planets and 63 known planetary satellites are shown in animated, to-scale drawings. The orbiting bodies move at their correct relative speeds about their parent, which is rendered as an attractive, to-scale gif image. Solar System Collisions: This most popular of our applications shows what happens when an asteroid or comet with user-defined size and speed impacts a given planet. The program calculates many effects, including the country impacted (if Earth is the target), energy of explosion, crater size, and magnitude of the ``planetquake'' generated. It also displays a relevant image (e.g. terrestrial crater, lunar crater, etc.). Scale of the Universe: Travel away from the Earth at a chosen speed and see how long it takes to reach other planets, stars and galaxies. This tool helps students visualize astronomical distances in an intuitive way. Scientific Notation: Students are interactively guided through conversions between scientific notation and regular numbers. Orbital Simulations: These tools allow the student to investigate different aspects of the three-body problem of celestial mechanics. Astronomy Workshop Bulletin Board: Get innovative teaching ideas and read about in-class experiences with the Astronomy Workshop. Share your ideas with other educators by posting on the Bulletin Board. Funding for the Astronomy Workshop is provided by NSF.

  14. The Dacum Workshop. Dacum 2.

    ERIC Educational Resources Information Center

    Mitchell, Barbara J.

    This booklet, the second in a series of four volumes on the Designing a Curriculum (Dacum) process, is intended to familiarize prospective Dacum workshop participants with the goals and procedures of the workshop. The first section of the booklet comprises a description of a Dacum workshop as a means of bringing together educators and employers to…

  15. Magnetic Suspension Technology Workshop

    NASA Technical Reports Server (NTRS)

    Keckler, Claude R. (Editor); Groom, Nelson J. (Editor); Britcher, Colin P. (Editor)

    1993-01-01

    In order to identify the state of magnetic suspension technology in such areas as rotating systems, pointing of experiments or subsystems, payload isolation, and superconducting materials, a workshop on Magnetic Suspension Technology was held at the Langley Research Center in Hampton, Virginia, on 2-4 Feb. 1988. The workshop included five technical sessions in which a total of 24 papers were presented. The technical sessions covered the areas of pointing, isolation, and measurement, rotating systems, modeling and control, and superconductors. A list of attendees is provided.

  16. Information Power Implementation Workshop.

    ERIC Educational Resources Information Center

    American Association of School Librarians, Chicago, IL.

    The materials in this collection were used at workshops designed to assist school library media specialists and learning resources center professionals in making effective use of "Information Power," a recent joint publication of the Association for Educational Communications and Technology (AECT) and the American Association of School Librarians…

  17. World without Workshops.

    ERIC Educational Resources Information Center

    Winkley, William M.

    1985-01-01

    The author examines the history of segregation for blind and handicapped persons and the relationship of segregation to social attitudes and opportunities. He also questions the relevance of sheltered workshops today. The El Paso Lighthouse "enclave with industry" program is presented as one model for moving toward a "natural proportion"…

  18. Flywheel energy storage workshop

    SciTech Connect

    O`Kain, D.; Carmack, J.

    1995-12-31

    Since the November 1993 Flywheel Workshop, there has been a major surge of interest in Flywheel Energy Storage. Numerous flywheel programs have been funded by the Advanced Research Projects Agency (ARPA), by the Department of Energy (DOE) through the Hybrid Vehicle Program, and by private investment. Several new prototype systems have been built and are being tested. The operational performance characteristics of flywheel energy storage are being recognized as attractive for a number of potential applications. Programs are underway to develop flywheels for cars, buses, boats, trains, satellites, and for electric utility applications such as power quality, uninterruptible power supplies, and load leveling. With the tremendous amount of flywheel activity during the last two years, this workshop should again provide an excellent opportunity for presentation of new information. This workshop is jointly sponsored by ARPA and DOE to provide a review of the status of current flywheel programs and to provide a forum for presentation of new flywheel technology. Technology areas of interest include flywheel applications, flywheel systems, design, materials, fabrication, assembly, safety & containment, ball bearings, magnetic bearings, motor/generators, power electronics, mounting systems, test procedures, and systems integration. Information from the workshop will help guide ARPA & DOE planning for future flywheel programs. This document is comprised of detailed viewgraphs.

  19. MOVES Workshops and Presentations

    EPA Pesticide Factsheets

    The U.S. EPA held a three-day workshop including EPA presentations on MOVES 2010 algorithms and default data, information on ways to use MOVES more efficiently for various purposes, and discussion of ideas and plans for MOVES future development.

  20. Microwave Workshop for Windows.

    ERIC Educational Resources Information Center

    White, Colin

    1998-01-01

    "Microwave Workshop for Windows" consists of three programs that act as teaching aid and provide a circuit design utility within the field of microwave engineering. The first program is a computer representation of a graphical design tool; the second is an accurate visual and analytical representation of a microwave test bench; the third…

  1. Dynamic defense workshop :

    SciTech Connect

    Crosby, Sean Michael; Doak, Justin E.; Haas, Jason Juedes.; Helinski, Ryan; Lamb, Christopher C.

    2013-02-01

    On September 5th and 6th, 2012, the Dynamic Defense Workshop: From Research to Practice brought together researchers from academia, industry, and Sandia with the goals of increasing collaboration between Sandia National Laboratories and external organizations, de ning and un- derstanding dynamic, or moving target, defense concepts and directions, and gaining a greater understanding of the state of the art for dynamic defense. Through the workshop, we broadened and re ned our de nition and understanding, identi ed new approaches to inherent challenges, and de ned principles of dynamic defense. Half of the workshop was devoted to presentations of current state-of-the-art work. Presentation topics included areas such as the failure of current defenses, threats, techniques, goals of dynamic defense, theory, foundations of dynamic defense, future directions and open research questions related to dynamic defense. The remainder of the workshop was discussion, which was broken down into sessions on de ning challenges, applications to host or mobile environments, applications to enterprise network environments, exploring research and operational taxonomies, and determining how to apply scienti c rigor to and investigating the eld of dynamic defense.

  2. African Outreach Workshop 1974.

    ERIC Educational Resources Information Center

    Schmidt, Nancy J.

    This report discusses the 1974 African Outreach Workshop planned and coordinated by the African Studies Program at the University of Illinois at Urbana-Champaign. Its major aim was to assist teachers in developing curriculum units on African using materials available in their local community. A second aim was for the African Studies Program to…

  3. Mentoring. Beginnings Workshop.

    ERIC Educational Resources Information Center

    Scallan-Berl, Patricia; Moguil, Leslie; Nyman, Sessy I.; Mercado, Miriam Mercado

    2003-01-01

    This workshop presents information on mentoring relationships within child care settings. Articles are: (1) "Mentoring Teachers...A Partnership in Learning" (Patricia Scallan-Berl); (2) "The Potential Gains of Peer Mentoring among Children" (Leslie Moguil); (3) "Mentoring Advocates in the Context of Early Childhood…

  4. Physics Teachers Workshop

    SciTech Connect

    Huggins, DaNel; Calhoun, John; Palmer, Alyson; Thorpe, Steve; Vanderveen, Anne

    2011-01-01

    INL is looking for the nation's top high school physics teachers to attend our July workshop in Idaho Falls. Participants get to learn from nuclear researchers, tour facilities including a research reactor and interact with peers from across the country. You can learn more about INL projects at http://www.facebook.com/idahonationallaboratory

  5. Child Nutrition. Beginnings Workshop.

    ERIC Educational Resources Information Center

    Hayden, Jacqueline; Eastman, Wayne; Aird, Laura Dutil; McCrea, Nadine L.

    2002-01-01

    Four workshops focus on nutrition for infants and children in child care settings. Articles are: (1) "Nutrition and Child Development: Global Perspectives" (Jacqueline Hayden); (2) "Working with Families around Nutritional Issues" (Wayne Eastman); (3) "Breastfeeding Promotion in Child Care" (Laura Dutil Aird); and (4) "Food as Shared…

  6. Polar Ozone Workshop. Abstracts

    NASA Technical Reports Server (NTRS)

    Aikin, Arthur C.

    1988-01-01

    Results of the proceedings of the Polar Ozone Workshop held in Snowmass, CO, on May 9 to 13, 1988 are given. Topics covered include ozone depletion, ozonometry, polar meteorology, polar stratospheric clouds, remote sensing of trace gases, atmospheric chemistry and dynamical simulations.

  7. Physics Teachers Workshop

    ScienceCinema

    Huggins, DaNel; Calhoun, John; Palmer, Alyson; Thorpe, Steve; Vanderveen, Anne

    2016-07-12

    INL is looking for the nation's top high school physics teachers to attend our July workshop in Idaho Falls. Participants get to learn from nuclear researchers, tour facilities including a research reactor and interact with peers from across the country. You can learn more about INL projects at http://www.facebook.com/idahonationallaboratory

  8. Synthetic Vision Workshop 2

    NASA Technical Reports Server (NTRS)

    Kramer, Lynda J. (Compiler)

    1999-01-01

    The second NASA sponsored Workshop on Synthetic/Enhanced Vision (S/EV) Display Systems was conducted January 27-29, 1998 at the NASA Langley Research Center. The purpose of this workshop was to provide a forum for interested parties to discuss topics in the Synthetic Vision (SV) element of the NASA Aviation Safety Program and to encourage those interested parties to participate in the development, prototyping, and implementation of S/EV systems that enhance aviation safety. The SV element addresses the potential safety benefits of synthetic/enhanced vision display systems for low-end general aviation aircraft, high-end general aviation aircraft (business jets), and commercial transports. Attendance at this workshop consisted of about 112 persons including representatives from industry, the FAA, and other government organizations (NOAA, NIMA, etc.). The workshop provided opportunities for interested individuals to give presentations on the state of the art in potentially applicable systems, as well as to discuss areas of research that might be considered for inclusion within the Synthetic Vision Element program to contribute to the reduction of the fatal aircraft accident rate. Panel discussions on topical areas such as databases, displays, certification issues, and sensors were conducted, with time allowed for audience participation.

  9. Pump Operation Workshop.

    ERIC Educational Resources Information Center

    Ontario Ministry of the Environment, Toronto.

    This manual was developed for use at workshops designed as an extension of training for water and wastewater treatment personnel. The course consists of lecture-discussions and hands-on activities. Each of the lessons in this document has clearly stated behavioral objectives to tell the trainee what he should know or do after completing that…

  10. Preventive Maintenance Workshop.

    ERIC Educational Resources Information Center

    Ontario Ministry of the Environment, Toronto.

    This manual was developed for use at workshops designed to upgrade the knowledge of experienced water and wastewater treatment plant operators. The course consists of lecture-discussions and hands-on activities. Each of the lessons has clearly stated behavioral objectives to tell the trainee what he should know or do after completing a topic.…

  11. Technology Leadership Workshop.

    ERIC Educational Resources Information Center

    Technology & Innovations in Education, Rapid City, SD.

    This Technology & Innovations in Education (TIE) workshop, presented in Kansas City, Missouri, on May 2, 1997, was designed to help participants gain a valid big picture of current school technology change issues, acquire current materials, clarify their beliefs, vision, and needs for their district's technology efforts, learn strategies for…

  12. Workshop on Cosmogenic Nuclides

    NASA Technical Reports Server (NTRS)

    Reedy, R. C. (Editor); Englert, P. (Editor)

    1986-01-01

    Abstracts of papers presented at the Workshop on Cosmogenic Nuclides are compiled. The major topic areas covered include: new techniques for measuring nuclides such as tandem accelerator and resonance mass spectrometry; solar modulation of cosmic rays; pre-irradiation histories of extraterrestrial materials; terrestrial studies; simulations and cross sections; nuclide production rate calculations; and meteoritic nuclides.

  13. Parent Conferences. Beginnings Workshop.

    ERIC Educational Resources Information Center

    Duffy, Roslyn; And Others

    1997-01-01

    Presents six workshop sessions on parent conferences: (1) "Parents' Perspectives on Conferencing" (R. Duffy); (2) "Three Way Conferences" (G. Zeller); (3) "Conferencing with Parents of Infants" (K. Albrecht); (4) "Conferencing with Parents of School-Agers" (L. G. Miller); (5) "Cross Cultural Conferences" (J. Gonzalez-Mena); and (6) "Working with…

  14. Fourth Airborne Geoscience Workshop

    NASA Technical Reports Server (NTRS)

    1991-01-01

    The focus of the workshop was on how the airborne community can assist in achieving the goals of the Global Change Research Program. The many activities that employ airborne platforms and sensors were discussed: platforms and instrument development; airborne oceanography; lidar research; SAR measurements; Doppler radar; laser measurements; cloud physics; airborne experiments; airborne microwave measurements; and airborne data collection.

  15. Workshop on Molecular Evolution

    NASA Technical Reports Server (NTRS)

    Cummings, Michael P.

    2004-01-01

    Molecular evolution has become the nexus of many areas of biological research. It both brings together and enriches such areas as biochemistry, molecular biology, microbiology, population genetics, systematics, developmental biology, genomics, bioinformatics, in vitro evolution, and molecular ecology. The Workshop provides an important contribution to these fields in that it promotes interdisciplinary research and interaction, and thus provides a glue that sticks together disparate fields. Due to the wide range of fields addressed by the study of molecular evolution, it is difficult to offer a comprehensive course in a university setting. It is rare for a single institution to maintain expertise in all necessary areas. In contrast, the Workshop is uniquely able to provide necessary breadth and depth by utilizing a large number of faculty with appropriate expertise. Furthermore, the flexible nature of the Workshop allows for rapid adaptation to changes in the dynamic field of molecular evolution. For example, the 2003 Workshop included recently emergent research areas of molecular evolution of development and genomics.

  16. [Cutaneous surgery workshop].

    PubMed

    Purim, Kátia Sheylla Malta

    2010-08-01

    The training of physician request knowledge, skills and attitudes for the effective exercise of professional practice. The training of basic surgical techniques, used in outpatient procedures, will prepare students to work in different scenarios. This work presents a proposal for teaching through workshops for cutaneous surgery in an experimental model.

  17. Radiation Source Replacement Workshop

    SciTech Connect

    Griffin, Jeffrey W.; Moran, Traci L.; Bond, Leonard J.

    2010-12-01

    This report summarizes a Radiation Source Replacement Workshop in Houston Texas on October 27-28, 2010, which provided a forum for industry and researchers to exchange information and to discuss the issues relating to replacement of AmBe, and potentially other isotope sources used in well logging.

  18. 76 FR 52954 - Workshop: Advancing Research on Mixtures; New Perspectives and Approaches for Predicting Adverse...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-24

    ... HUMAN SERVICES Workshop: Advancing Research on Mixtures; New Perspectives and Approaches for Predicting... ``Advancing Research on Mixtures: New Perspectives and Approaches for Predicting Adverse Human Health Effects....niehs.nih.gov/conferences/dert/mixtures/ . The deadline to register for this workshop is...

  19. Immunity to Polyomavirus BK Infection: Immune Monitoring to Regulate the Balance between Risk of BKV Nephropathy and Induction of Alloimmunity

    PubMed Central

    Cioni, Michela; Basso, Sabrina; Gagliardone, Chiara; Potenza, Leonardo; Verrina, Enrico; Luppi, Mario; Zecca, Marco; Ghiggeri, Gian Marco; Ginevri, Fabrizio

    2013-01-01

    Polyomavirus BK-associated nephropathy (PyVAN) is the main infectious cause of allograft damage after kidney transplantation. A number of studies revealed an association between the presence of BKV-specific cellular immunity and BK viral clearance, with patients failing to recover specific T cells progressing to PyVAN. Evolution to allograft dysfunction can be prevented by restoration of BKV-specific immunity through a stepwise reduction of maintenance immunosuppressive drugs. Prospective monitoring of BK viral load and specific immunity, together with B-cell alloimmune surveillance, may allow a targeted modification/reduction of immunosuppression, with the aim of obtaining viral clearance while preventing graft injury due to deposition of de novo donor-specific HLA antibodies and late/chronic antibody-mediated allograft injury. Innovative, immune-based therapies may further contribute to BKV infection prevention and control. PMID:24000288

  20. AASTRA Leadership Workshop

    NASA Astrophysics Data System (ADS)

    Hemenway, M. K.

    1998-05-01

    American Astronomical Society Teacher Resource Agent Institutes were held in the summers of 1994-1996. From the 215 Agents, sixteen were selected to attend a follow-up leadership workshop for three weeks in July, 1997 at University of Texas at Austin. The workshop followed the recommendations of both the AASTRA formative evaluator and the National Science Education Standards in allowing the participants time to reflect on their practice as teachers and as teacher-leaders. At no financial charge to the project, the Southwest Educational Development Laboratory contributed three workshops of three hours each. These workshops, plus one given by the PI, were taken from "Facilitating Systemic Change in Science and Mathematics Education : A Toolkit for Professional Developers" [ISBN 1-878234-08-0]. In addition, McDonald Observatory contributed six nights of telescope time on the 30-inch telescope with a CCD prime focus camera. In Austin, several astronomers lectured on their research; most of the lectures correlated with the research projects in progress at the Observatory during the teachers' observing run. Several observers allowed the teachers to participate in their observing sessions on the larger telescopes. Participant evaluation of the Leadership Workshop was very positive, both in terms of enhancing their self-image as leaders, enriching their repertoire of activities to use within their classrooms, and introducing them to modern astronomical research techniques. AASTRA is supported by NSF under grant ESI 93- 53377 and the AAS. Additional support is provided by Loyola University of Chicago, Northern Arizona University of Flagstaff, University of Maryland at College Park, and the University of Texas at Austin.

  1. Merkel Cell Carcinoma: A Virus-Induced Human Cancer

    PubMed Central

    Chang, Yuan; Moore, Patrick S.

    2013-01-01

    Merkel cell polyomavirus (MCV) is the first polyomavirus directly linked to human cancer, and its recent discovery helps to explain many of the enigmatic features of Merkel cell carcinoma (MCC). MCV is clonally integrated into MCC tumor cells, which then require continued MCV oncoprotein expression to survive. The integrated viral genomes have a tumor-specific pattern of tumor antigen gene mutation that incapacitates viral DNA replication. This human cancer virus provides a new model in which a common, mostly harmless member of the human viral flora can initiate cancer if it acquires a precise set of mutations in a host with specific susceptibility factors, such as age and immune suppression. Identification of this tumor virus has led to new opportunities for early diagnosis and targeted treatment of MCC. PMID:21942528

  2. BrainMap `95 workshop

    SciTech Connect

    1995-12-31

    The fourth annual BrainMap workshop was held at La Mansion del Rio Hotel in San Antonio December 3--4, 1995. The conference title was ``Human Brain Mapping and Modeling.`` The meeting was attended by 137 registered participants and 30 observers from 82 institutions representing 12 countries. The meeting focused on the technical issues associated with brain mapping and modeling. A total of 23 papers were presented covering the following topics: spatial normalization and registration; functional image analysis; metanalysis and modeling; and new horizons in biological databases. The full program with abstracts was available on the Research Imaging Center`s web site. A book will be published by John Wiley and Sons prior to the end of 1998.

  3. International workshop of chromosome 19

    SciTech Connect

    Pericak-Vance, M.A. . Div. of Neurology); Carrano, A.J. )

    1991-09-16

    This document summarizes the workshop on physical and genetic mapping of chromosome 19. The first session discussed the major disease loci found on the chromosome. The second session concentrated on reference families, markers and linkage maps. The third session concentrated on radiation hybrid mapping, somatic cell hybrid panels, macro restriction maps and YACs, followed by cDNA and long range physical maps. The fourth session concentrated on compiling consensus genetic and physical maps as well as discussing regions of conflict. The final session dealt with the LLNL cosmid contig database and comparative mapping of homologous regions of the human and mouse genomes, and ended with a discussion of resource sharing. 18 refs., 2 figs. (MHB)

  4. UVI Cyber-security Workshop Workshop Analysis.

    SciTech Connect

    Kuykendall, Tommie G.; Allsop, Jacob Lee; Anderson, Benjamin Robert; Boumedine, Marc; Carter, Cedric; Galvin, Seanmichael Yurko; Gonzalez, Oscar; Lee, Wellington K.; Lin, Han Wei; Morris, Tyler Jake; Nauer, Kevin S.; Potts, Beth A.; Ta, Kim Thanh; Trasti, Jennifer; White, David R.

    2015-07-08

    The cybersecurity consortium, which was established by DOE/NNSA’s Minority Serving Institutions Partnerships Program (MSIPP), allows students from any of the partner schools (13 HBCUs, two national laboratories, and a public school district) to have all consortia options available to them, to create career paths and to open doors to DOE sites and facilities to student members of the consortium. As a part of this year consortium activities, Sandia National Laboratories and the University of Virgin Islands conducted a week long cyber workshop that consisted of three courses; Digital Forensics and Malware Analysis, Python Programming, and ThunderBird Cup. These courses are designed to enhance cyber defense skills and promote learning within STEM related fields.

  5. The Affording Mars Workshop: Background and Recommendations

    NASA Technical Reports Server (NTRS)

    Thronson, Harley A.; Carberry, Christopher

    2014-01-01

    A human mission to Mars is the stated "ultimate" goal for NASA and is widely believed by the public to be the most compelling destination for America's space program. However, widely cited enormous costs - perhaps as much as a trillion dollars for a many-decade campaign - seem to be an impossible hurdle, although political and budget instability over many years may be equally challenging. More recently, a handful of increasingly detailed architectures for initial Mars missions have been developed by commercial companies that have estimated costs much less than widely believed and roughly comparable with previous major human space flight programs: the Apollo Program, the International Space Station, and the space shuttle. Several of these studies are listed in the bibliography to the workshop report. As a consequence of these new scenarios, beginning in spring, 2013 a multiinstitutional planning team began developing the content and invitee list for a winter workshop that would critically assess concepts, initiatives, technology priorities, and programmatic options to reduce significantly the costs of human exploration of Mars. The output of the workshop - findings and recommendations - would be presented in a number of forums and discussed with national leaders in human space flight. It would also be made available to potential international partners. This workshop was planned from the start to be the first in a series. Subsequent meetings, conferences, and symposia will concentrate on topics not able to be covered in December. In addition, to make progress in short meeting, a handful of ground rules were adopted by the planning team and agreed to by the participants. Perhaps the two most notable such ground rules were (1) the Space Launch System (SLS) and Orion would be available during the time frame considered by the participants and (2) the International Space Station (ISS) would remain the early linchpin in preparing for Mars exploration over the coming decade

  6. New Perspectives on Human Problem Solving

    ERIC Educational Resources Information Center

    Goldstone, Robert L.; Pizlo, Zygmunt

    2009-01-01

    In November 2008 at Purdue University, the 2nd Workshop on Human Problem Solving was held. This workshop, which was a natural continuation of the first workshop devoted almost exclusively to optimization problems, addressed a wider range of topics that reflect the scope of the "Journal of Problem Solving." The workshop was attended by 35…

  7. New findings about trichodysplasia spinulosa-associated polyomavirus (TSPyV)--novel qPCR detects TSPyV-DNA in blood samples.

    PubMed

    Urbano, Paulo R; Nali, Luiz H S; Bicalho, Camila S; Pierrotti, Ligia C; David-Neto, Elias; Pannuti, Cláudio S; Romano, Camila M

    2016-02-01

    A new real-time PCR assay for trichodysplasia spinulosa-associated polyomavirus (TSPyV) DNA detection was designed, and blood samples from kidney transplant recipients and healthy individuals were screened. TSPyV-DNA was not detected in blood from healthy individuals, but 26.8% of kidney recipients presented TSPyV-DNA. This is the first report of TSPyV viremia.

  8. Composites Damage Tolerance Workshop

    NASA Technical Reports Server (NTRS)

    Gregg, Wayne

    2006-01-01

    The Composite Damage Tolerance Workshop included participants from NASA, academia, and private industry. The objectives of the workshop were to begin dialogue in order to establish a working group within the Agency, create awareness of damage tolerance requirements for Constellation, and discuss potential composite hardware for the Crew Launch Vehicle (CLV) Upper Stage (US) and Crew Module. It was proposed that a composites damage tolerance working group be created that acts within the framework of the existing NASA Fracture Control Methodology Panel. The working group charter would be to identify damage tolerance gaps and obstacles for implementation of composite structures into manned space flight systems and to develop strategies and recommendations to overcome these obstacles.

  9. Geoengineering design parameters workshop

    SciTech Connect

    St. John, C.M. and Associates, Grand Junction, CO ); Kim, Kunsoo . Rockwell Hanford Operations)

    1985-12-12

    A one-day workshop on the subject of the geotechnical design parameters, in situ stress and rock mass strength, for a nuclear waste repository in basalt was held in Rapid City, South Dakota, on June 25, 1989. A panel comprised of five widely recognized experts in the field of rock mechanics, met to discuss the state of stress at the Hanford Site and the strength of a basalt rock mass. This report summarizes the discussions that took place and presents a set of final position statements developed collaboratively by the panel and the workshop moderator. The report concludes with a set of specific recommendations for future actions considered necessary to adequately define the in situ stress and the rock mass strength at the Hanford Site and to document the position of the Basalt Waste Isolation Project in respect to these two critical design parameters.

  10. Imaging Sciences Workshop Proceedings

    SciTech Connect

    Candy, J.V.

    1996-11-21

    This report contains the proceedings of the Imaging Sciences Workshop sponsored by C.A.S.LS., the Center for Advanced Signal & Image Sciences. The Center, established primarily to provide a forum where researchers can freely exchange ideas on the signal and image sciences in a comfortable intellectual environment, has grown over the last two years with the opening of a Reference Library (located in Building 272). The Technical Program for the 1996 Workshop include a variety of efforts in the Imaging Sciences including applications in the Microwave Imaging, highlighted by the Micro-Impulse Radar (MIR) system invented at LLNL, as well as other applications in this area. Special sessions organized by various individuals in Speech, Acoustic Ocean Imaging, Radar Ocean Imaging, Ultrasonic Imaging, and Optical Imaging discuss various applica- tions of real world problems. For the more theoretical, sessions on Imaging Algorithms and Computed Tomography were organized as well as for the more pragmatic featuring a session on Imaging Systems.

  11. Mars Surface Mission Workshop

    NASA Technical Reports Server (NTRS)

    Duke, M. B. (Editor)

    1997-01-01

    A workshop was held at the Lunar and Planetary Institute on September 4-5, 1997, to address the surface elements of the Mars Reference Mission now being reviewed by NASA. The workshop considered the current reference mission and addressed the types of activities that would be expected for science and resource exploration and facilities operations. A set of activities was defined that can be used to construct "vignettes" of the surface mission. These vignettes can form the basis for describing the importance of the surface mission, for illustrating aspects of the surface mission, and for allowing others to extend and revise these initial ideas. The topic is rich with opportunities for additional conceptualization. It is recommended that NASA consider supporting university design teams to conduct further analysis of the possibilities.

  12. Summary of the workshop

    SciTech Connect

    Ruggiero, A.G.

    1988-01-01

    The Workshop on the ''RHIC'' Performance was held at the Collider Center, Brookhaven National Laboratory, from March 21 to 26, 1988. The Workshop had three major goals: To check the validity and soundness of the methods described in the Conceptual Design Report (CDR, May 1986), in order to ensure the performance proposed there. To initiate and study techniques that increase the luminosity in the second stage of RHIC by a factor of twenty beyond the CDR values, and to reduce the interactive region to a rms length of less than 20 cm. To investigate possible limits on beam intensity and dimensions, and their impact on RHIC hardware. Conclusions and recommendations on these topics are discussed in this paper. 5 tabs.

  13. Actinide Spectroscopy Workshop

    SciTech Connect

    Tobin, J.G.; Shuh, D.K.

    2004-12-05

    Actinide materials present an extreme scientific challenge to the materials research community. The complex electronic structures of actinide materials result in many unusual and unique properties that have yet to be fully understood. The difficulties in handling, preparing, and characterizing actinide materials has frequently precluded investigations and has the limited the detailed understanding of these relevant, complex materials. However, modern experiments with actinide materials have the potential to provide key, fundamental information about many long-standing issues concerning actinide materials. This workshop focused on the scientific and technical challenges posed by actinide materials and the potential that synchrotron radiation approaches available at the ALS can contribute to improving the fundamental understanding of actinides materials. Fundamental experimental approaches and results, as well as theoretical modeling and computational simulations, were part of the workshop program.

  14. Rayleigh Scattering Diagnostics Workshop

    NASA Technical Reports Server (NTRS)

    Seasholtz, Richard (Compiler)

    1996-01-01

    The Rayleigh Scattering Diagnostics Workshop was held July 25-26, 1995 at the NASA Lewis Research Center in Cleveland, Ohio. The purpose of the workshop was to foster timely exchange of information and expertise acquired by researchers and users of laser based Rayleigh scattering diagnostics for aerospace flow facilities and other applications. This Conference Publication includes the 12 technical presentations and transcriptions of the two panel discussions. The first panel was made up of 'users' of optical diagnostics, mainly in aerospace test facilities, and its purpose was to assess areas of potential applications of Rayleigh scattering diagnostics. The second panel was made up of active researchers in Rayleigh scattering diagnostics, and its purpose was to discuss the direction of future work.

  15. Magnet measurement workshop

    SciTech Connect

    1986-12-01

    This report covers the deliberations of the participants the workshop and some subsequent contributions. Section III, the report of the rotating coil group, includes a summary table of the major measuring systems in use today, with separate sections on each. Section IV is the summary report of the group that addressed other measuring techniques. Because one of the limits of all the techniques being considered is electronic data acquisition, Section V addresses this topic. A set of issues relevant to magnetic field measurements of SSC dipoles was raised and addressed during the workshop. These are included as Section VI. Section VII includes a complete list of attendees with their addresses and a separate list of the members of the two working groups.

  16. PREFACE: Collapse Calderas Workshop

    NASA Astrophysics Data System (ADS)

    Gottsmann, Jo; Aguirre-Diaz, Gerardo

    2008-10-01

    Caldera-formation is one of the most awe-inspiring and powerful displays of nature's force. Resultant deposits may cover vast areas and significantly alter the immediate topography. Post-collapse activity may include resurgence, unrest, intra-caldera volcanism and potentially the start of a new magmatic cycle, perhaps eventually leading to renewed collapse. Since volcanoes and their eruptions are the surface manifestation of magmatic processes, calderas provide key insights into the generation and evolution of large-volume silicic magma bodies in the Earth's crust. Despite their potentially ferocious nature, calderas play a crucial role in modern society's life. Collapse calderas host essential economic deposits and supply power for many via the exploitation of geothermal reservoirs, and thus receive considerable scientific, economic and industrial attention. Calderas also attract millions of visitors world-wide with their spectacular scenic displays. To build on the outcomes of the 2005 calderas workshop in Tenerife (Spain) and to assess the most recent advances on caldera research, a follow-up meeting was proposed to be held in Mexico in 2008. This abstract volume presents contributions to the 2nd Calderas Workshop held at Hotel Misión La Muralla, Querétaro, Mexico, 19-25 October 2008. The title of the workshop `Reconstructing the evolution of collapse calderas: Magma storage, mobilisation and eruption' set the theme for five days of presentations and discussions, both at the venue as well as during visits to the surrounding calderas of Amealco, Amazcala and Huichapan. The multi-disciplinary workshop was attended by more than 40 scientist from North, Central and South America, Europe, Australia and Asia. Contributions covered five thematic topics: geology, geochemistry/petrology, structural analysis/modelling, geophysics, and hazards. The workshop was generously supported by the International Association of Volcanology and the Chemistry of The Earth's Interior

  17. Modified Composite Materials Workshop

    NASA Technical Reports Server (NTRS)

    Dicus, D. L. (Compiler)

    1978-01-01

    The reduction or elimination of the hazard which results from accidental release of graphite fibers from composite materials was studied at a workshop. At the workshop, groups were organized to consider six topics: epoxy modifications, epoxy replacement, fiber modifications, fiber coatings and new fibers, hybrids, and fiber release testing. Because of the time required to develop a new material and acquire a design data base, most of the workers concluded that a modified composite material would require about four to five years of development and testing before it could be applied to aircraft structures. The hybrid working group considered that some hybrid composites which reduce the risk of accidental fiber release might be put into service over the near term. The fiber release testing working group recommended a coordinated effort to define a suitable laboratory test.

  18. Interdisciplinary workshop in the philosophy of medicine: medical knowledge, medical duties

    PubMed Central

    Kingma, Elselijn

    2014-01-01

    Abstract On 27 September 2013, the Centre for the Humanities and Health (CHH) at King's College London hosted a 1‐day workshop on ‘Medical knowledge, Medical Duties’. This workshop was the fifth in a series of five workshops whose aim is to provide a new model for high‐quality, open interdisciplinary engagement between medical professionals and philosophers. This report identifies the key points of discussion raised throughout the day and the methodology employed. PMID:25470528

  19. Research needs to better understand Lake Ontario ecosystem function: A workshop summary

    USGS Publications Warehouse

    Stewart, Thomas J.; Rudstam, Lars G.; Watkins, James M.; Johnson, Timothy B.; Weidel, Brian C.; Koops, Marten A.

    2016-01-01

    Lake Ontario investigators discussed and interpreted published and unpublished information during two workshops to assess our current understanding of Lake Ontario ecosystem function and to identify research needs to guide future research and monitoring activities. The purpose of this commentary is to summarize key investigative themes and hypotheses that emerged from the workshops. The outcomes of the workshop discussions are organized under four themes: spatial linkages and interactions, drivers of primary production, trophic transfer, and human interactions.

  20. Interdisciplinary workshop in the philosophy of medicine: medical knowledge, medical duties.

    PubMed

    Bullock, Emma; Kingma, Elselijn

    2014-12-01

    On 27 September 2013, the Centre for the Humanities and Health (CHH) at King's College London hosted a 1-day workshop on 'Medical knowledge, Medical Duties'. This workshop was the fifth in a series of five workshops whose aim is to provide a new model for high-quality, open interdisciplinary engagement between medical professionals and philosophers. This report identifies the key points of discussion raised throughout the day and the methodology employed.