The muscles of the athletes to learn surface anatomy - The Influence of classical statues on anatomy teaching

Gross anatomy classes are still regarded as an integral part of human biomedical education worldwide. The first documentary evidence of the practice of anatomical dissection for teaching purposes dates back to the 13th century AD, although this practice seems to have originated in Ancient Greece, if not in earlier times. Dissection of the human body is practiced in most anatomy institutions worldwide despite increasing pressure to reduce material and staff costs, regardless the ongoing debate concerning the suitability of body donors for medical education. Moreover, anatomical teaching skills are also evolving and need to be tailored for the different areas of anatomical expertise students have to acquire: therefore, anatomic dissection goes probably beyond the scope of anatomy teaching in some classes such as sports sciences. However, there is no doubt that a practical approach to the study and teaching of anatomy is surely preferable to basic ex cathedra anatomy lectures. Here, we propose a new teaching method for sports sciences and fine arts students by training their surface anatomy skills through the study of ancient statues

SARS-CoV-2 diagnostics in the virology laboratory of a University Hospital in Rome during the lockdown period

Italy was one of the most affected nations by coronavirus disease 2019 outside China. The infections, initially limited to Northern Italy, spread to all other Italian regions. This study aims to provide a snapshot of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) epidemiology based on a single-center laboratory experience in Rome. The study retrospectively included 6565 subjects tested for SARS-CoV-2 at the Laboratory of Virology of Sapienza University Hospital in Rome from 6 March to 4 May. A total of 9995 clinical specimens were analyzed, including nasopharyngeal swabs, bronchoalveolar lavage fluids, gargle lavages, stools, pleural fluids, and cerebrospinal fluids. Positivity to SARS-CoV-2 was detected in 8% (527/6565) of individuals, increased with age, and was higher in male patients (P <.001). The number of new confirmed cases reached a peak on 18 March and then decreased. The virus was detected in respiratory samples, in stool and in pleural fluids, while none of gargle lavage or cerebrospinal fluid samples gave a positive result. This analysis allowed to gather comprehensive information on SARS-CoV-2 epidemiology in our area, highlighting positivity variations over time and in different sex and age group and the need for a continuous surveillance of the infection, mostly because the pandemic evolution remains unknown

Merkel cell polyomavirus (MCPyV) in the context of Immunosuppression. Genetic analysis of noncoding control region (NCCR) variability among a HIV-1-positive population

Background: Since limited data are available about the prevalence of Merkel cell polyomavirus (MCPyV) and the genetic variability of its noncoding control region (NCCR) in the context of immunosuppression, this study aimed to investigate the distribution of MCPyV in anatomical sites other than the skin and the behavior of NCCR among an HIV-1-positive population. Methods: Urine, plasma, and rectal swabs specimens from a cohort of 66 HIV-1-positive patients were collected and subjected to quantitative real-time polymerase chain reaction (qPCR) for MCPyV DNA detection. MCPyV-positive samples were amplified by nested PCR targeting the NCCR, and NCCRs alignment was carried out to evaluate the occurrence of mutations and to identify putative binding sites for cellular factors. Results: MCPyV DNA was detected in 10/66 urine, in 7/66 plasma, and in 23/66 rectal samples, with a median value of 5 × 102 copies/mL, 1.5 × 102 copies/mL, and 2.3 × 103 copies/mL, respectively. NCCR sequence analysis revealed a high degree of homology with the MCC350 reference strain in urine, whereas transitions, transversions, and single or double deletions were observed in plasma and rectal swabs. In these latter samples, representative GTT and GTTGA insertions were also observed. Search for putative binding sites of cellular transcription factors showed that in several strains, deletions, insertions, or single base substitutions altered the NCCR canonical configuration. Conclusions: Sequencing analysis revealed the presence of numerous mutations in the NCCR, including insertions and deletions. Whether these mutations may have an impact on the pathogenic features of the virus remains to be determined. qPCR measured on average a low viral load in the specimens analyzed, with the exception of those with the GTTGA insertion

Fracionamento de carbono em nitossolo e cambissolo com aplicação de fertilizantes organominerais.

O teor de carbono orgânico do solo pode sofrer alterações com as práticas agrícolas adotadas em especial, devido ao uso de fertilizantes orgânicos. O objetivo foi quantificar os teores e estoques de carbono orgânico total (COT), particulado (COP), associado aos minerais (COam), em Nitossolo e Cambissolo submetidos à aplicação de fertilizantes minerais e organominerais nas formas sólidas e fluidas. Os tratamentos caracterizam fatorial 2x5, em blocos casualizados com grupos de experimento e quatro repetições, descritos a seguir: fator A solo - Nitossolo Vermelho eutroférrico típico e Cambissolo Háplico eutroférrico léptico; fator B adubação, sendo: controle (C), organomineral fluído (OF), organomineral sólido (OS), mineral fluído (MF) e mineral sólido (MS). A resposta foi avaliada após cultivos sucessivos de milho e aveia, sorgo forrageiro e trigo, no período de 2010 a 2013. Após três anos de plantio direto com as culturas de milho, sorgo, aveia e trigo com adoção da prática agrícola de adubação com diferentes fertilizantes organominerais e minerais nas formas sólidas e fluidas, poucas alterações nas diferentes frações de C orgânico no solo foram constatadas, com significância apenas no tratamento MF para Cambissolo em COT e COam na camada de 0-5 e 0-20 cm e COP na camada de 5-10cm, resultados que refletem no estoque de C em COT e COam na camada de 0-20cm onde MF foi igual a MS e OS e estes superiores aos demais tratamentos. Verificou-se diferença para índice de manejo do solo (IMC) apenas na camada 5-10 cm no Cambissolo, sendo o MF igual ao OS e superior aos demais tratamentos, o que coloca estes tratamentos em evidência para esta classe de solo e profundidade específica

Involvement of the Chloroplastic Isoform of tRNA Ligase in the Replication of Viroids Belonging to the Family Avsunviroidae

Avocado sunblotch viroid, peach latent mosaic viroid, chrysanthemum chlorotic mottle viroid, and eggplant latent viroid (ELVd), the four recognized members of the family Avsunviroidae, replicate through the symmetric pathway of an RNA-to-RNA rolling-circle mechanism in chloroplasts of infected cells. Viroid oligomeric transcripts of both polarities contain embedded hammerhead ribozymes that, during replication, mediate their self-cleavage to monomeric-length RNAs with 5'-hydroxyl and 2',3'-phosphodiester termini that are subsequently circularized. We report that a recombinant version of the chloroplastic isoform of the tRNA ligase from eggplant (Solanum melongena L.) efficiently catalyzes in vitro circularization of the plus [(+)] and minus [(-)] monomeric linear replication intermediates from the four Avsunviroidae. We also show that while this RNA ligase specifically recognizes the genuine monomeric linear (+) ELVd replication intermediate, it does not do so with five other monomeric linear (+) ELVd RNAs with their ends mapping at different sites along the molecule, despite containing the same 5'-hydroxyl and 2',3'-phosphodiester terminal groups. Moreover, experiments involving transient expression of a dimeric (+) ELVd transcript in Nicotiana benthamiana Domin plants preinoculated with a tobacco rattle virus-derived vector to induce silencing of the plant endogenous tRNA ligase show a significant reduction of ELVd circularization. In contrast, circularization of a viroid replicating in the nucleus occurring through a different pathway is unaffected. Together, these results support the conclusion that the chloroplastic isoform of the plant tRNA ligase is the host enzyme mediating circularization of both (+) and (-) monomeric linear intermediates during replication of the viroids belonging to the family Avsunviroidae.This work was supported by the Ministerio de Ciencia e Innovacion (MICINN) from Spain through grants BIO2008-01986, BIO2011-26741, and BFU2008-03154. M. A. Nohales and D. Molina-Serrano were the recipients of predoctoral fellowships from the Spanish Ministerio de Educacion y Ciencia.Nohales Zafra, MA.; Molina Serrano, D.; Flores Pedauye, R.; Daros Arnau, JA. (2012). Involvement of the Chloroplastic Isoform of tRNA Ligase in the Replication of Viroids Belonging to the Family Avsunviroidae. Journal of Virology. 86:8269-8276. https://doi.org/10.1128/JVI.00629-12S8269827686Abelson, J., Trotta, C. R., & Li, H. (1998). tRNA Splicing. 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Peach latent mosaic viroid is locked by a 2′,5′-phosphodiester bond produced by in vitro self-ligation 1 1Edited by D. E. Draper. Journal of Molecular Biology, 273(3), 533-543. doi:10.1006/jmbi.1997.1355Daros, J.-A. (2002). A chloroplast protein binds a viroid RNA in vivo and facilitates its hammerhead-mediated self-cleavage. The EMBO Journal, 21(4), 749-759. doi:10.1093/emboj/21.4.749Daros, J. A., Marcos, J. F., Hernandez, C., & Flores, R. (1994). Replication of avocado sunblotch viroid: evidence for a symmetric pathway with two rolling circles and hammerhead ribozyme processing. Proceedings of the National Academy of Sciences, 91(26), 12813-12817. doi:10.1073/pnas.91.26.12813De la Pena, M. (2003). Peripheral regions of natural hammerhead ribozymes greatly increase their self-cleavage activity. The EMBO Journal, 22(20), 5561-5570. doi:10.1093/emboj/cdg530De la Pena, M., Navarro, B., & Flores, R. (1999). Mapping the molecular determinant of pathogenicity in a hammerhead viroid: A tetraloop within the in vivo branched RNA conformation. Proceedings of the National Academy of Sciences, 96(17), 9960-9965. doi:10.1073/pnas.96.17.9960Ding, B. (2009). The Biology of Viroid-Host Interactions. Annual Review of Phytopathology, 47(1), 105-131. doi:10.1146/annurev-phyto-080508-081927Englert, M. (2005). Plant tRNA ligases are multifunctional enzymes that have diverged in sequence and substrate specificity from RNA ligases of other phylogenetic origins. Nucleic Acids Research, 33(1), 388-399. doi:10.1093/nar/gki174Englert, M., Latz, A., Becker, D., Gimple, O., Beier, H., & Akama, K. (2007). Plant pre-tRNA splicing enzymes are targeted to multiple cellular compartments. Biochimie, 89(11), 1351-1365. doi:10.1016/j.biochi.2007.06.014Fadda, Z., Daros, J. A., Fagoaga, C., Flores, R., & Duran-Vila, N. (2003). Eggplant Latent Viroid, the Candidate Type Species for a New Genus within the Family Avsunviroidae (Hammerhead Viroids). Journal of Virology, 77(11), 6528-6532. doi:10.1128/jvi.77.11.6528-6532.2003Feldstein, P. A., Hu, Y., & Owens, R. A. (1998). Precisely full length, circularizable, complementary RNA: An infectious form of potato spindle tuber viroid. Proceedings of the National Academy of Sciences, 95(11), 6560-6565. doi:10.1073/pnas.95.11.6560Flores, R., Daròs, J.-A., & Hernández, C. (2000). Avsunviroidae family: Viroids containing hammerhead ribozymes. Advances in Virus Research, 271-323. doi:10.1016/s0065-3527(00)55006-4Flores, R., Hernández, C., Alba, A. E. M. de, Daròs, J.-A., & Serio, F. D. (2005). Viroids and Viroid-Host Interactions. Annual Review of Phytopathology, 43(1), 117-139. doi:10.1146/annurev.phyto.43.040204.140243Flores, R., & Owens, R. A. (2008). Viroids. Encyclopedia of Virology, 332-342. doi:10.1016/b978-012374410-4.00532-xGas, M.-E., Hernández, C., Flores, R., & Daròs, J.-A. (2007). Processing of Nuclear Viroids In Vivo: An Interplay between RNA Conformations. PLoS Pathogens, 3(11), e182. doi:10.1371/journal.ppat.0030182Gas, M.-E., Molina-Serrano, D., Hernandez, C., Flores, R., & Daros, J.-A. (2008). Monomeric Linear RNA of Citrus Exocortis Viroid Resulting from Processing In Vivo Has 5’-Phosphomonoester and 3’-Hydroxyl Termini: Implications for the RNase and RNA Ligase Involved in Replication. Journal of Virology, 82(20), 10321-10325. doi:10.1128/jvi.01229-08Gómez, G., & Pallás, V. (2010). Noncoding RNA Mediated Traffic of Foreign mRNA into Chloroplasts Reveals a Novel Signaling Mechanism in Plants. PLoS ONE, 5(8), e12269. doi:10.1371/journal.pone.0012269Hernandez, C., & Flores, R. (1992). Plus and minus RNAs of peach latent mosaic viroid self-cleave in vitro via hammerhead structures. Proceedings of the National Academy of Sciences, 89(9), 3711-3715. doi:10.1073/pnas.89.9.3711Hertel, K. J., Herschlag, D., & Uhlenbeck, O. C. (1994). A Kinetic and Thermodynamic Framework for the Hammerhead Ribozyme Reaction. Biochemistry, 33(11), 3374-3385. doi:10.1021/bi00177a031Hutchins, C. J., Keese, P., Visvader, J. E., Rathjen, P. D., McInnes, J. L., & Symons, R. H. (1985). Comparison of multimeric plus and minus forms of viroids and virusoids. Plant Molecular Biology, 4(5), 293-304. doi:10.1007/bf02418248Hutchins, C. J., Rathjen, P. D., Forster, A. C., & Symons, R. H. (1986). Self-cleavage of plus and minus RNA transcripts of avocado sunblotch viroid. Nucleic Acids Research, 14(9), 3627-3640. doi:10.1093/nar/14.9.3627Khvorova, A., Lescoute, A., Westhof, E., & Jayasena, S. D. (2003). Sequence elements outside the hammerhead ribozyme catalytic core enable intracellular activity. Nature Structural & Molecular Biology, 10(9), 708-712. doi:10.1038/nsb959Kiberstis, P. A., Haseloff, J., & Zimmern, D. (1985). 2′ phosphomonoester, 3′-5′ phosphodiester bond at a unique site in a circular viral RNA. The EMBO Journal, 4(3), 817-822. doi:10.1002/j.1460-2075.1985.tb03703.xKonarska, M., Filipowicz, W., Domdey, H., & Gross, H. J. (1981). Formation of a 2′-phosphomonoester, 3′,5′-phosphodiester linkage by a novel RNA ligase in wheat germ. Nature, 293(5828), 112-116. doi:10.1038/293112a0Liu, Y., Schiff, M., Marathe, R., & Dinesh-Kumar, S. P. (2002). Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. The Plant Journal, 30(4), 415-429. doi:10.1046/j.1365-313x.2002.01297.xMakino, S., Sawasaki, T., Endo, Y., & Takai, K. (2005). Purification and sequence determination of an RNA ligase from wheat embryos. Nucleic Acids Symposium Series, 49(1), 319-320. doi:10.1093/nass/49.1.319Marcos, J. F., & Flores, R. (1993). The 5’ end Generated in the in vitro Self-Cleavage Reaction of Avocado Sunblotch Viroid RNAs is Present in Naturally Occurring Linear Viroid Molecules. Journal of General Virology, 74(5), 907-910. doi:10.1099/0022-1317-74-5-907Martinez, F., Marques, J., Salvador, M. L., & Daros, J.-A. (2009). Mutational analysis of eggplant latent viroid RNA processing in Chlamydomonas reinhardtii chloroplast. Journal of General Virology, 90(12), 3057-3065. doi:10.1099/vir.0.013425-0Molina-Serrano, D., Marqués, J., Nohales, M.-Á., Flores, R., & Daròs, J.-A. (2012). A chloroplastic RNA ligase activity analogous to the bacterial and archaeal 2´–5′ RNA ligase. RNA Biology, 9(3), 326-333. doi:10.4161/rna.19218Navarro, B., & Flores, R. (1997). Chrysanthemum chlorotic mottle viroid: Unusual structural properties of a subgroup of self-cleaving viroids with hammerhead ribozymes. Proceedings of the National Academy of Sciences, 94(21), 11262-11267. doi:10.1073/pnas.94.21.11262Navarro, J.-A., Daròs, J.-A., & Flores, R. (1999). Complexes Containing Both Polarity Strands of Avocado Sunblotch Viroid: Identification in Chloroplasts and Characterization. Virology, 253(1), 77-85. doi:10.1006/viro.1998.9497Navarro, J.-A., Vera, A., & Flores, R. (2000). A Chloroplastic RNA Polymerase Resistant to Tagetitoxin Is Involved in Replication of Avocado Sunblotch Viroid. Virology, 268(1), 218-225. doi:10.1006/viro.1999.0161Nelson, J. A., Shepotinovskaya, I., & Uhlenbeck, O. C. (2005). Hammerheads Derived from sTRSV Show Enhanced Cleavage and Ligation Rate Constants†. Biochemistry, 44(44), 14577-14585. doi:10.1021/bi051130tPRODY, G. A., BAKOS, J. T., BUZAYAN, J. M., SCHNEIDER, I. R., & BRUENING, G. (1986). Autolytic Processing of Dimeric Plant Virus Satellite RNA. 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Analisi di scenario per la riduzione delle emissioni inquinanti nella Regione Puglia

Nello scenario energetico ed ambientale nazionale, la definizione di metodologie innovative per la valutazione delle emissioni inquinanti in atmosfera, appare di notevole importanza al fine di operare nell’ambito di politiche energetiche ottimali, sia per la necessità di rispettare gli obiettivi di emissione di CO2 previsti dal Protocollo di Kyoto e dall’Unione Europea, sia per il soddisfacimento del crescente fabbisogno energetico senza un ulteriore uso di combustibili fossili. Il presente lavoro ha come obiettivo quello di individuare il potenziale di riduzione delle emissioni inquinanti nella Regione Puglia, utilizzando tecnologie che sfruttano le fonti rinnovabili di energia (impianti fotovoltaici, mini impianti eolici) e tecniche avanzate di risparmio energetico (impianti di co-trigenerazione con microturbine a gas naturale). Utilizzando il modello Long-range Energy Alternatives Planning System (LEAP) è stato possibile individuare quali politiche ambientali sostenere e, conseguentemente, sui quali processi di cambiamento dei comportamenti individuali e collettivi intervenire, al fine di condurre il sistema verso gli obiettivi ambientali programmati e misurare gli effetti delle scelte politiche sull’ambiente

COS-7-based model: methodological approach to study John Cunningham virus replication cycle

John Cunningham virus (JCV) is a human neurotropic polyomavirus whose replication in the Central Nervous System (SNC) induces the fatal demyelinating disease, progressive multifocal leukoencephalopathy (PML). JCV propagation and PML investigation have been severely hampered by the lack of an animal model and cell culture systems to propagate JCV have been very limited in their availability and robustness. We previously confirmed that JCV CY strain efficiently replicated in COS-7 cells as demonstrated by the progressive increase of viral load by quantitative PCR (Q-PCR) during the time of transfection and that archetypal regulatory structure was maintained, although two characteristic point mutations were detected during the viral cycle. This short report is an important extension of our previous efforts in defining our reliable model culture system able to support a productive JCV infection. Supernatants collected from transfected cells have been used to infect freshly seeded COS-7 cell line. An infectious viral progeny was obtained as confirmed by Western blot and immunofluorescence assay. During infection, the archetype regulatory region was conserved. Importantly, in this study we developed an improved culture system to obtain a large scale production of JC virus in order to study the genetic features, the biology and the pathogenic mechanisms of JC virus that induce PML

COS-7-based model: methodological approach to study John Cunningham virus replication cycle

John Cunningham virus (JCV) is a human neurotropic polyomavirus whose replication in the Central Nervous System (SNC) induces the fatal demyelinating disease, progressive multifocal leukoencephalopathy (PML). JCV propagation and PML investigation have been severely hampered by the lack of an animal model and cell culture systems to propagate JCV have been very limited in their availability and robustness. We previously confirmed that JCV CY strain efficiently replicated in COS-7 cells as demonstrated by the progressive increase of viral load by quantitative PCR (Q-PCR) during the time of transfection and that archetypal regulatory structure was maintained, although two characteristic point mutations were detected during the viral cycle. This short report is an important extension of our previous efforts in defining our reliable model culture system able to support a productive JCV infection. Supernatants collected from transfected cells have been used to infect freshly seeded COS-7 cell line. An infectious viral progeny was obtained as confirmed by Western blot and immunofluorescence assay. During infection, the archetype regulatory region was conserved. Importantly, in this study we developed an improved culture system to obtain a large scale production of JC virus in order to study the genetic features, the biology and the pathogenic mechanisms of JC virus that induce PML