Characterization of the promoter region of ftsZ from Corynebacterium glutamicum and controlled overexpression of FtsZ

Of the five promoters detected for the ftsZ gene in Corynebacterium glutamicum, three were located within the coding region of the upstream ftsQ gene and two within the intergenic ftsQ-ftsZ region. The most distant ftsZ promoter showed activity in Escherichia coli and controlled high-level transcriptional expression of ftsZ in C. glutamicum. Quantitative Western blotting showed that all five promoters were active during the exponential growth phase and down-regulated during stationary phase. This tightly controlled expression of ftsZ in C. glutamicum indicated that small changes in the amount of FtsZ protein strongly affect bacterial cell viability. The controlled overexpression of ftsZ in C. glutamicum, using the promoter of the gntK gene (PgntK), resulted in approximately 5-fold overproduction of FtsZ, an increase in cell diameter, and a highly variable localization of the protein as spirals or tangles throughout the cell. These results suggest that the intracellular concentration of FtsZ is critical for productive septum formation in C. glutamicum. Our observations provide insight into the mechanisms used by the coryneform group, which lacks actin homologs and many regulators of cell division, to control cell morphology. [Int Microbiol 2007; 10(4): 271-282

Sobre verbos denominales: construcciones causativas y de localización

Los verbos denominales están formados a partir de una base sustantiva. Estos verbos pueden expresar diferentes tipos de contenido semántico; en este artículo nos centramos en aquellos que manifiestan significado causativo y local. Concretamente, se analizan los verbos denominales causativos y los verbos denominales locales y se demuestra la necesidad de diferenciar un subgrupo de verbos denominales causativo-localizadores en los que es tan relevante el contenido causativo como el local. En estos verbos, la figura que se localiza no es previa a la localización, como sucede en el caso de los esquemas puramente locales, sino que surge como resultado de la acción causativa. Por lo tanto, estos verbos expresan localización y causatividad de forma intrínseca y no de forma circunstancial.We form denominal causative verbs from nouns. Although these verbs can express different kinds of meaning, we will focus here on causative and local meanings. Specifically, we will analyze denominal causative verbs and denominal local verbs, showing, thus, the need to differentiate a subgroup of denominal causative-local verbs, where causative meaning proves to be as important as local meaning. In these verbs, the localised figure doesn’t show prior to location, as it happens in the case of typical local schemas, but emerges as a result of the causative action. Therefore, denominal causative-local verbs express location and causativity inherently and not accidentally.Este trabajo ha sido posible gracias al proyecto de investigación HUM2006-09429/FILO, financiado por el M.E.C

Utilización de antibióticos en una farmacia comunitaria

La sociedad actual se caracteriza por una elevada intolerancia al malestar y una urgencia por solucionar molestias, incluso menores, que conduce a una demanda de utilización de los recursos disponibles, aunque no siempre se precisen, lo que influye en que el uso, entre otros, de antibióticos sea mucho mayor de lo deseable

Impact of SARS-CoV-2 Infection on Patients with Cancer: Retrospective and Transversal Studies in Spanish Population

[EN] Background: Studies of patients with cancer affected by coronavirus disease 2019 (COVID-19) are needed to assess the impact of the disease in this sensitive population, and the influence of different cancer treatments on the COVID-19 infection and seroconversion. Material and Methods: We performed a retrospective analysis of all patients hospitalized with RT-PCR positive for COVID-19 in our region to assess the prevalence of cancer patients and describe their characteristics and evolution (Cohort 1). Concurrently, a transversal study was carried out in patients on active systemic cancer treatment for symptomatology and seroprevalence (IgG/IgM by ELISA-method) against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) (Cohort 2). Results: A total of 215 patients (Cohort 1) were admitted to hospital with a confirmed COVID-19 infection between February 28 and April 30, 2020, and 17 died (7.9%). A medical record of cancer was noted in 43 cases (20%), 6 of them required Intensive care unit ICU attention (14%), and 7 died (16%). There were thirty-six patients (83%) who tested IgG/IgM positive for SARS-CoV-2. Patients on immunosuppressive therapies presented a lower ratio of seroconversion (40% vs. 8%; p = 0.02). In Cohort 2, 166 patients were included in a symptoms-survey and tested for SARS-CoV-2. Any type of potential COVID-19-related symptom was referred up to 67.4% of patients (85.9% vs. 48.2% vs. 73.9%, for patients on chemotherapy, immunotherapy and targeted therapies respectively, p < 0.05). The seroprevalence ratio was 1.8% for the whole cohort with no significant differences by patient or treatment characteristics. Conclusion: Patients with cancer present higher risks for hospital needs for COVID-19 infection. The lack of SARS-CoV-2 seroconversion may be a concern for patients on immunosuppressive therapies. Patients receiving systematic therapies relayed a high rate of potentially COVID-19-related symptoms, particularly those receiving chemotherapy. However, the seroconversion rate remains low and in the range of general population.We thank all the patients who consented to this study, and the frontline healthcare professionals who are involved in patients' care during this pandemic. We also thank the technical assistants: M. Portero Hernandez, A. Real Perez, and M. Ocasar Garcia. VGB's research work is partially supported by the Ministerio de Ciencia e Innovacion of Spain under grant No. PID2019-110442GB-I00.Garde-Noguera, J.; Fernández-Murga, ML.; Giner-Bosch, V.; Domínguez-Márquez, V.; García Sánchez, J.; Soler-Cataluña, JJ.; López Chuliá, F.... (2020). Impact of SARS-CoV-2 Infection on Patients with Cancer: Retrospective and Transversal Studies in Spanish Population. Cancers. 12(12):1-11. https://doi.org/10.3390/cancers12123513S1111212Munster, V. J., Koopmans, M., van Doremalen, N., van Riel, D., & de Wit, E. (2020). A Novel Coronavirus Emerging in China — Key Questions for Impact Assessment. New England Journal of Medicine, 382(8), 692-694. doi:10.1056/nejmp2000929Coronavirus COVID-19 Global Cases by the Center for Systems Science and Engineeringhttps://coronavirus.jhu.edu/map.htmlInforme Sobre la Situación COVID-19 en España a 6 April 2020. Equipo COVID-19. RENAVE. CNE. CNM (ISCIII). Informe COVID-19 nº21https://covid19.isciii.es/Report 13: Estimating the Number of Infections and the Impact of Non-Pharmaceutical Interventions on COVID-19 in 11 European Countrieshttps://spiral.imperial.ac.uk:8443/handle/10044/1/77731Rothan, H. A., & Byrareddy, S. N. (2020). The epidemiology and pathogenesis of coronavirus disease (COVID-19) outbreak. Journal of Autoimmunity, 109, 102433. doi:10.1016/j.jaut.2020.102433Liang, W., Guan, W., Chen, R., Wang, W., Li, J., Xu, K., … He, J. (2020). Cancer patients in SARS-CoV-2 infection: a nationwide analysis in China. The Lancet Oncology, 21(3), 335-337. doi:10.1016/s1470-2045(20)30096-6Sharpe, A. H., & Pauken, K. E. (2017). The diverse functions of the PD1 inhibitory pathway. Nature Reviews Immunology, 18(3), 153-167. doi:10.1038/nri.2017.108Fisher, R. A. (1922). On the Interpretation of χ 2 from Contingency Tables, and the Calculation of P. Journal of the Royal Statistical Society, 85(1), 87. doi:10.2307/2340521Mehta, C. R., & Patel, N. R. (1983). A Network Algorithm for Performing Fisher’s Exact Test in r × c Contingency Tables. Journal of the American Statistical Association, 78(382), 427. doi:10.2307/2288652Wilcoxon, F. (1945). Individual Comparisons by Ranking Methods. Biometrics Bulletin, 1(6), 80. doi:10.2307/3001968Kuderer, N. M., Choueiri, T. K., Shah, D. P., Shyr, Y., Rubinstein, S. M., Rivera, D. R., … de Lima Lopes, G. (2020). Clinical impact of COVID-19 on patients with cancer (CCC19): a cohort study. The Lancet, 395(10241), 1907-1918. doi:10.1016/s0140-6736(20)31187-9Zhang, L., Zhu, F., Xie, L., Wang, C., Wang, J., Chen, R., … Zhou, M. (2020). Clinical characteristics of COVID-19-infected cancer patients: a retrospective case study in three hospitals within Wuhan, China. Annals of Oncology, 31(7), 894-901. doi:10.1016/j.annonc.2020.03.296Winter, A. K., & Hegde, S. T. (2020). The important role of serology for COVID-19 control. The Lancet Infectious Diseases, 20(7), 758-759. doi:10.1016/s1473-3099(20)30322-4Venkatesulu, B. P., Thoguluva Chandrasekar, V., Giridhar, P., Advani, P., Sharma, A., Hsieh, C. E., … Krishnan, S. (2020). A systematic review and meta-analysis of cancer patients affected by a novel coronavirus. doi:10.1101/2020.05.27.20115303Van Assen, S., Holvast, A., Benne, C. A., Posthumus, M. D., van Leeuwen, M. A., Voskuyl, A. E., … Bijl, M. (2010). Humoral responses after influenza vaccination are severely reduced in patients with rheumatoid arthritis treated with rituximab. Arthritis & Rheumatism, 62(1), 75-81. doi:10.1002/art.25033Rousseau, B., Loulergue, P., Mir, O., Krivine, A., Kotti, S., Viel, E., … Tournigand, C. (2012). Immunogenicity and safety of the influenza A H1N1v 2009 vaccine in cancer patients treated with cytotoxic chemotherapy and/or targeted therapy: the VACANCE study. Annals of Oncology, 23(2), 450-457. doi:10.1093/annonc/mdr141Di Cosimo, S., Malfettone, A., Pérez-García, J. M., Llombart-Cussac, A., Miceli, R., Curigliano, G., & Cortés, J. (2020). Immune checkpoint inhibitors: a physiology-driven approach to the treatment of coronavirus disease 2019. European Journal of Cancer, 135, 62-65. doi:10.1016/j.ejca.2020.05.026Bersanelli, M., Scala, S., Affanni, P., Veronesi, L., Colucci, M. E., Banna, G. L., … Liotta, F. (2020). Immunological insights on influenza infection and vaccination during immune checkpoint blockade in cancer patients. Immunotherapy, 12(2), 105-110. doi:10.2217/imt-2019-0200Pollán, M., Pérez-Gómez, B., Pastor-Barriuso, R., Oteo, J., Hernán, M. A., Pérez-Olmeda, M., … Fernández de Larrea, N. (2020). Prevalence of SARS-CoV-2 in Spain (ENE-COVID): a nationwide, population-based seroepidemiological study. The Lancet, 396(10250), 535-544. doi:10.1016/s0140-6736(20)31483-5Choe, P. G., Perera, R. A. P. M., Park, W. B., Song, K.-H., Bang, J. H., Kim, E. S., … Oh, M. (2017). MERS-CoV Antibody Responses 1 Year after Symptom Onset, South Korea, 2015. Emerging Infectious Diseases, 23(7), 1079-1084. doi:10.3201/eid2307.170310Cao, W.-C., Liu, W., Zhang, P.-H., Zhang, F., & Richardus, J. H. (2007). Disappearance of Antibodies to SARS-Associated Coronavirus after Recovery. New England Journal of Medicine, 357(11), 1162-1163. doi:10.1056/nejmc070348Wu, L.-P., Wang, N.-C., Chang, Y.-H., Tian, X.-Y., Na, D.-Y., Zhang, L.-Y., … Liang, G.-D. (2007). Duration of Antibody Responses after Severe Acute Respiratory Syndrome. Emerging Infectious Diseases, 13(10), 1562-1564. doi:10.3201/eid1310.070576Kissler, S. M., Tedijanto, C., Goldstein, E., Grad, Y. H., & Lipsitch, M. (2020). Projecting the transmission dynamics of SARS-CoV-2 through the postpandemic period. Science, 368(6493), 860-868. doi:10.1126/science.abb579

Human Coronavirus Virulence Motifs and Virulence

Trabajo presentado en el XIV International Nidovirus Symposium (Nido2017), celebrado en Kansas City, Missouri (Estados Unidos), del 4 al 9 de junio de 2017We have shown that SARS-CoV E protein is a virulence factor that includes at least two virulence motifs: its ion channel (IC) activity encoded within the transmembrane domain and a PDZ binding motif (PBM) located at its carboxy-terminus. We showed that E protein pathogenicity was caused by the activation of different host signaling pathways. One of them was the activation of inflammasome, a process mediated by the conductance of Ca++ byEprotein IC activity, leading to an increased expression of IL-1beta, TNF-alpha and IL-6 levels. Another signaling pathway implied the activation of a proinflammatory response mediated by NF-kB activation. This activation was a consequence of E protein-syntenin binding mediated by PBM-PDZ interactions. This binding caused an increase of p38MAPK phosphorylation promoting the induction of acute respiratory distress syndrome (ARDS), edema and death of mice infected with a mouse adapted SARS-CoV. The relevance of p38 MAPK activation after infection with the mouse adapted SARS-CoV was confirmed by the protection of mice in the presence of an inhibitor of p38 MAPK, but not in its absence. These results illustrated the identification of an efficient coronavirus (CoV) antiviral. The presence of a virulence factor such as the PBM motif in E protein allows the virus to interact with more than 400 cell proteins containing PDZ motifs, conferring the virus the potential to control a high number of cell-signaling pathways increasing its replication and virulence. In fact, we are analyzing the proteome of the viral PBM-cellular PDZ interactions using system biology approaches. Frequently, the ARDS caused by lung infection with mild respiratory viruses is resolved before it evolves to serious edema. In contrast, after SARS-CoV infection frequently this resolution does not take place. We have shown the binding of E protein to a main mediator of edema resolution, the Na+ /K+ ATPase, and proposed that this may be one of the procedures by which edema recovery is prevented after SARS-CoV infection, either by inhibition of Na+ /K+ ATPase activity or by relocating this enzyme to another subcellular compartment. Deadly human CoVs as SARS- and MERS-CoVs have at least two viral proteins with IC activity and PBM motifs. Studies on the relevance of E and 3a SARS-CoV proteins in replication and virulence, and the interdependence among them have shown that the presence in the virus of at least E or 3a proteins was needed for virus viability. In fact, we have shown that the complementation between E and 3a proteins is mediated by the PBM motifs located at the carboxy-terminus of these proteins. Our studies on the interaction of SARS-CoV and MERS-CoV with the host, and the engineering of reverse genetics systems for each of these viruses, led us to the development of genetically stable vaccine candidates that provided full-protection against the challenge with the homologous virulent virus using mice models

Nature of viruses and pandemics: Coronaviruses

Coronaviruses (CoVs) have the largest genome among RNA viruses and store large amounts of information without genome integration as they replicate in the cell cytoplasm. The replication of the virus is a continuous process, whereas the transcription of the subgenomic mRNAs is a discontinuous one, involving a template switch, which resembles a high frequency recombination mechanism that may favor virus genome variability. The origin of the three deadly human CoVs SARS-CoV, MERS-CoV and SARS-CoV-2 are zoonotic events. SARS-CoV-2 has incorporated in its spike protein a furine proteolytic site that facilitates the activation of the virus in any tissue, making this CoV strain highly polytropic and pathogenic. Using MERS-CoV as a model, a propagation-deficient RNA replicon was generated by removing E protein gene (essential for viral morphogenesis and involved in virulence), and accessory genes 3, 4a, 4b and 5 (responsible for antagonism of the innate immune response) to attenuate the virus: MERS-CoV-Δ[3,4a,4b,5,E]. This RNA replicon is strongly attenuated and elicits sterilizing protection after a single immunization in transgenic mice with the receptor for MERS-CoV, making it a promising vaccine candidate for this virus and an interesting platform for vector-based vaccine development. A strategy could be developed for the design of RNA replicon vaccines for other human pathogenic coronaviruses.This work was supported by grants from the Government of Spain (PID2019-107001RB-I00 AEI/FEDER, UE; SEV 2017-0712 and PIE_INTRAMURAL_LINEA 1-202020E079), the CSIC (PIE_INTRAMURAL-202020E043), the European Commission (ISOLDA_848166 H2020-SC1-2019-Two-Stage-RTD, RIA; MANCO_101003651 H2020-SC1-PHE-CORONAVIRUS-2020 RIA), and the U.S. National Institutes of Health (NIH_2P01AI060699).Peer reviewe

SARS-CoV-2-Mediated Lung Edema and Replication Are Diminished by Cystic Fibrosis Transmembrane Conductance Regulator Modulators

20 Pág.Coronaviruses (CoVs) of genera α, β, γ, and δ encode proteins that have a PDZ-binding motif (PBM) consisting of the last four residues of the envelope (E) protein (PBM core). PBMs may bind over 400 cellular proteins containing PDZ domains (an acronym formed by the combination of the first letter of the names of the three first proteins where this domain was identified), making them relevant for the control of cell function. Three highly pathogenic human CoVs have been identified to date: severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV), and SARS-CoV-2. The PBMs of the three CoVs were virulence factors. SARS-CoV mutants in which the E protein PBM core was replaced by the E protein PBM core from virulent or attenuated CoVs were constructed. These mutants showed a gradient of virulence, depending on whether the alternative PBM core introduced was derived from a virulent or an attenuated CoV. Gene expression patterns in the lungs of mice infected with SARS-CoVs encoding each of the different PBMs were analyzed by RNA sequencing of infected lung tissues. E protein PBM of SARS-CoV and SARS-CoV-2 dysregulated gene expression related to ion transport and cell homeostasis. Decreased expression of cystic fibrosis transmembrane conductance regulator (CFTR) mRNA, essential for alveolar edema resolution, was shown. Reduced CFTR mRNA levels were associated with edema accumulation in the alveoli of mice infected with SARS-CoV and SARS-CoV-2. Compounds that increased CFTR expression and activity, significantly reduced SARS-CoV-2 growth in cultured cells and protected against mouse infection, suggesting that E protein virulence is mediated by a decreased CFTR expression. IMPORTANCE Three highly pathogenic human CoVs have been identified: SARS-CoV, MERS-CoV, and SARS-CoV-2. The E protein PBMs of these three CoVs were virulence factors. Gene expression patterns associated with the different PBM motifs in the lungs of infected mice were analyzed by deep sequencing. E protein PBM motif of SARS-CoV and SARS-CoV-2 dysregulated the expression of genes related to ion transport and cell homeostasis. A decrease in the mRNA expression of the cystic fibrosis transmembrane conductance regulator (CFTR), which is essential for edema resolution, was observed. The reduction of CFTR mRNA levels was associated with edema accumulation in the lungs of mice infected with SARS-CoV-2. Compounds that increased the expression and activity of CFTR drastically reduced the production of SARS-CoV-2 and protected against its infection in a mice model. These results allowed the identification of cellular targets for the selection of antivirals.This work was supported by grants from the Government of Spain (BIO2016-75549-R; PID2019-107001RB-I00 AEI/FEDER, UE; SEV 2017-0712 and PIE_INTRAMURAL_LINEA 1- 202020E079), CSIC (PIE_INTRAMURAL-202020E043), the European Zoonotic Anticipation and Preparedness Initiative (ZAPI) (IMI_JU_115760), the European Commission (H2020-SC1- 2019, ISOLDA Project No. 848166-2), and the U.S. National Institutes of Health (NIH) (2P01AI060699). J.M.H. received a contract from Comunidad de Madrid (Y2020/BIO-6576, COVID-PREclinical-MODels-CM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. In vivo experiments were performed at INIA-CISA (Madrid, Spain)Peer reviewe

Las vacunas del CNB-CSIC contra la COVID-19: La vacuna basada en un coronavirus inofensivo

Cómic realizado en el marco del proyecto de divulgación "Ciencia y Responsabilidad Frente a la COVID19", financiado dentro de la convocatoria Cuenta la Ciencia de la Fundación General del CSIC (FGCSIC) e impulsado por Susanna Manrubia, investigadora del CNB-CSIC.-- El cómic ha sido realizado por Jesús Sánchez Ruiz (Laboratoons) y ha contado con la supervisión científica del laboratorio de Luis Enjuanes e Isabel Sola (a través de José Honrubia, Alejandro Sanz y Javier Gutiérrez).Colección completa: 1. ¿Podemos predecir el comportamiento de la población durante una pandemia? http://hdl.handle.net/10261/244009.-- 2. ¡Cómo combatir un apocalipsis zombi! http://hdl.handle.net/10261/244014.-- 3. Las vacunas del CNB-CSIC contra la COVID-19: La vacuna basada en un coronavirus inofensivo. http://hdl.handle.net/10261/244019.-- 4. Las vacunas del CNB-CSIC contra la COVID-19: La vacuna basada en el virus vaccinia. http://hdl.handle.net/10261/244023.-- 5.-- ¿Cómo trabajamos con algo que no vemos? http://hdl.handle.net/10261/244024. 6. ¿Podemos defendernos de un virus extraterrestre? http://hdl.handle.net/10261/244028Peer reviewe

Edema macular quístico por toxoplasma gondii

We study a case of a 17 years old woman that were visited by urgency because of an ocular pain and sudden visual loss (VA) on right eye 4 days ago. At diagnostic moment she had a VA of 0.2 and a moderate inflammatory activity on anterior chamber. On fundoscope study she presented a serous macular detachment with a paramacular whitish lesion and littlies haemorrhages. Angiographic study revealed a foveal detachment of neurosensorial retina with late hyperfluorescence. We confirmed diagnostic by serology. After treatment the patient was better gradually. Actually she has a good VA and a inactivity scar on paramacular area. The most common manifestation of toxoplasmosis is a retinochoroiditis. Ocular toxoplasmic diagnostic can be doing by funduscopy findings and exclusion diagnostic. Cystoid macular edema occurs when lesions are close to the macular area. We can confirm diagnostic by serology. Adequate treatment are pyrimetamine, sulfadiacine and systemic steroids.Presentamos el caso de una paciente de 17 años vista de urgencias por dolor y disminución de agudeza visual en ojo derecho de 4 días de evolución. En el momento del diagnóstico presentaba una AV de 0.2 y una actividad inflamatoria en cámara anterior moderada. En el estudio fundoscópico presentaba un desprendimiento seroso macular acompañado de una lesión blanquecina paramacular y pequeñas hemorragias adyacentes. La angiografía reveló un desprendimiento foveal de la retina neurosensorial con hiperfluorescencia franca en las fases tardías. Se confirma el diagnóstico de toxoplasmosis mediante serología. Tras la instauración de tratamiento el cuadro mejora progresivamente hasta alcanzar una AV de la unidad y un resto cicatricial parafoveal. La manifestación ocular más frecuente de una toxoplasmosis es la coriorretinitis. El diagnóstico de toxoplasmosis ocular puede hacerse tras la observación de las lesiones típicas en fondo de ojo y descartando otras causas de coriorretinitis. Podemos confirmar el diagnóstico por serología. El edema macular cistoide se produce cuando la lesión está cerca de la mácula. El tratamiento adecuado combina la triada pirimetamina, sulfadiazina y esteroides sistémicos