A new challenge for meteorological measurements: The meteoMet project-Metrology for meteorology

Climate change and its consequences require immediate actions in order to safeguard the environment and economy in Europe and in the rest of world. Aiming to enhance data reliability and reduce uncertainties in climate observations, a joint research project called MeteoMet-Metrology for Meteorology started in October 2011 coordinated by the Italian Istituto Nazionale di Ricerca Metrologica (INRiM). The project is focused on the traceability of measurements involved in climate change: surface and upper air measurements of temperature, pressure, humidity, wind speed and direction, solar irradiance and reciprocal influences between measurands. This project will provide the first definition at the European level of validated climate parameters with associated uncertainty budgets and novel criteria for interpretation of historical data series. The big challenge is the propagation of a metrological measurement perspective to meteorological observations. When such an approach will be adopted the requirement of reliable data and robust datasets over wide scales and long terms could be better met. © 2013 AIP Publishing LLC

MDM2 and Fbw7 cooperate to induce p63 protein degradation following DNA damage and cell differentiation

Tight control of p63 protein levels must be achieved under differentiation or apoptotic conditions. Here, we describe a new regulatory pathway for the \u394Np63\u3b1 protein. We found that MDM2 binds \u394Np63\u3b1 in the nucleus promoting its translocation to the cytoplasm. The MDM2 nuclear localization signal is required for \u394Np63\u3b1 nuclear export and subsequent degradation, whereas the MDM2 ring-finger domain is dispensable. Once exported to the cytoplasm by MDM2, p63 is targeted for degradation by the Fbw7 E3-ubiquitin ligase. Efficient degradation of \u394Np63\u3b1 by Fbw7 (also known as FBXW7) requires GSK3 kinase activity. By deletion and point mutations analysis we have identified a phosphodegron located in the \u3b1 and \u3b2 tail of p63 that is required for degradation. Furthermore, we show that MDM2 or Fbw7 depletion inhibits degradation of endogenous \u394Np63\u3b1 in cells exposed to UV irradiation, adriamycin and upon keratinocyte differentiation. Our findings suggest that following DNA damage and cellular differentiation MDM2 and Fbw7 can cooperate to regulate the levels of the pro-proliferative \u394Np63\u3b1 protein

Capability of national reference laboratories in Latin America to detect emerging resistance mechanisms

Fil: Corso, Alejandra. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Guerriero, Leonor. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Pasterán, Fernando. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Ceriana, Paola. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Callejo, Raquel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Bacteriología Especial; Argentina.Fil: Prieto, Mónica. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Bacteriología Especial; Argentina.Fil: Tuduri, Ezequiel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Lopardo, Horacio. Hospital de Pediatría S.A.M.I.C. "Prof Dr. Juan Garrahan"; Argentina.Fil: Vay, Carlos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Hospital de Clínicas "José de San Martín"; Argentina.Fil: Smayevsky, Jorgelina. Centro de Educación Médica e Investigaciones Clínicas "Norberto Quirno"; Argentina.Fil: Tokumoto, Marta. Fundación Favaloro; Argentina.Fil: Alvarez, Jorge Matheu. Organización Panamericana de la Salud. Área de Vigilancia de la Salud y Prevención y Control de Enfermedades; Estados Unidos.Fil: Ramón Pardo, Pilar. Organización Panamericana de la Salud. Área de Vigilancia de la Salud y Prevención y Control de Enfermedades; Estados Unidos.Galas, Marcelo. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.OBJETIVO: Evaluar la capacidad de 17 laboratorios nacionales de referencia que participan en el Programa Latinoamericano de Control de Calidad en Bacteriología y Resistencia a los Antimicrobianos (LA-EQAS) para detectar mecanismos de resistencia emergentes, a saber: resistencia de enterobacterias a carbapenemes por presencia de Klebsiella pneumoniae carbapenemasa (KPC); resistencia de enterobacterias a carbapenemes por presencia de metalobetalactamasas (MBL) tipo IMP, y resistencia intermedia a vancomicina de aislamientos de Staphylococcus aureus (VISA). MÉTODOS: Se enviaron los siguientes tres aislamientos a los 17 laboratorios participantes del LA-EQAS: Klebsiella pneumoniae OPS-161 productor de KPC, Enterobacter cloacae OPS-166 productor de IMP y S. aureus OPS-165 con resistencia intermedia a vancomicina. Se evaluó la interpretación de las pruebas de sensibilidad y detección del mecanismo de resistencia y el tamaño de los halos de inhibición (método de difusión por discos) o valor de la concentración inhibitoria mínima (CIM). RESULTADOS: La concordancia en la detección de los mecanismos de resistencia fue de 76,4%, 73,3% y 66,7% con respecto a la cepas K. pneumoniae OPS-161, E. cloacae OPS-166 y S. aureus OPS-165, respectivamente. La concordancia entre las zonas de inhibición obtenidas por los laboratorios participantes y los rangos establecidos por el laboratorio coordinador fue aceptable en los tres aislamientos, ya que alcanzó 90,8%, 92,8% y 88,9%, respectivamente, para cada cepa. CONCLUSIONES: La concordancia global en la detección de los mecanismos de resistencia KPC, MBL y VISA fue de 72,1%. Consideramos que los laboratorios nacionales de referencia de América Latina son capaces de reconocer estos mecanismos de resistencia emergentes y se espera que en el futuro la concordancia alcance su nivel máximo

Emergence of Respiratory Streptococcus agalactiae Isolates in Cystic Fibrosis Patients

Streptococcus agalactiae is a well-known pathogen for neonates and immunocompromized adults. Beyond the neonatal period, S. agalactiae is rarely found in the respiratory tract. During 2002–2008 we noticed S. agalactiae in respiratory secretions of 30/185 (16%) of cystic fibrosis (CF) patients. The median age of these patients was 3–6 years older than the median age CF patients not harboring S. agalactiae. To analyze, if the S. agalactiae isolates from CF patients were clonal, further characterization of the strains was achieved by capsular serotyping, surface protein determination and multilocus sequence typing (MLST). We found a variety of sequence types (ST) among the isolates, which did not substantially differ from the MLST patterns of colonizing strains from Germany. However serotype III, which is often seen in colonizing strains and invasive infections was rare among CF patients. The emergence of S. agalactiae in the respiratory tract of CF patients may represent the adaptation to a novel host environment, supported by the altered surfactant composition in older CF patients

Claudin-1 Is a p63 Target Gene with a Crucial Role in Epithelial Development

The epidermis of the skin is a self-renewing, stratified epithelium that functions as the interface between the human body and the outer environment, and acts as a barrier to water loss. Components of intercellular junctions, such as Claudins, are critical to maintain tissue integrity and water retention. p63 is a transcription factor essential for proliferation of stem cells and for stratification in epithelia, mutated in human hereditary syndromes characterized by ectodermal dysplasia. Both p63 and Claudin-1 null mice die within few hours from birth due to dehydration from severe skin abnormalities. These observations suggested the possibility that these two genes might be linked in one regulatory pathway with p63 possibly regulating Claudin-1 expression. Here we show that silencing of ΔNp63 in primary mouse keratinocytes results in a marked down-regulation of Claudin-1 expression (−80%). ΔNp63α binds in vivo to the Claudin-1 promoter and activates both the endogenous Claudin-1 gene and a reporter vector containing a –1.4 Kb promoter fragment of the Claudin-1 gene. Accordingly, Claudin-1 expression was absent in the skin of E15.5 p63 null mice and natural p63 mutant proteins, specifically those found in Ankyloblepharon–Ectodermal dysplasia–Clefting (AEC) patients, were indeed altered in their capacity to regulate Claudin-1 transcription. This correlates with deficient Claudin-1 expression in the epidermis of an AEC patient carrying the I537T p63 mutation. Notably, AEC patients display skin fragility similar to what observed in the epidermis of Claudin-1 and p63 null mice. These findings reinforce the hypothesis that these two genes might be linked in a common regulatory pathway and that Claudin-1 may is an important p63 target gene involved in the pathogenesis of ectodermal dysplasias

Analysis of Large Phenotypic Variability of EEC and SHFM4 Syndromes Caused by K193E Mutation of the TP63 Gene

EEC (ectrodactyly, ectodermal dysplasia, clefting; OMIM 604292) is an autosomal dominant developmental disorder resulting mainly from pathogenic mutations of the DNA-binding domain (DBD) of the TP63 gene. In this study, we showed that K193E mutation in nine affected individuals of a four-generation kindred with a large degree of phenotypic variability causes four different syndromes or TP63-related disorders: EEC, Ectrodactyly-ectodermal dysplasia (EE), isolated ectodermal dysplasia, and isolated Split Hand/Foot Malformation type 4 (SHFM4). Genotype-phenotype and DBD structural modeling analysis showed that the K193-located loop L2-A is associated with R280 through hydrogen bonding interactions, while R280 mutations also often cause large phenotypic variability of EEC and SHFM4. Thus, we speculate that K193 and several other DBD mutation-associated syndromes may share similar pathogenic mechanisms, particularly in the case of the same mutation with different phenotypes. Our study and others also suggest that the phenotypic variability of EEC is attributed, at least partially, to genetic and/or epigenetic modifiers

Molecular, microbiological and clinical characterization of Clostridium difficile isolates from tertiary care hospitals in Colombia

In Colombia, the epidemiology and circulating genotypes of Clostridium difficile have not yet been described. Therefore, we molecularly characterized clinical isolates of C.difficile from patients with suspicion of C.difficile infection (CDI) in three tertiary care hospitals. C.difficile was isolated from stool samples by culture, the presence of A/B toxins were detected by enzyme immunoassay, cytotoxicity was tested by cell culture and the antimicrobial susceptibility determined. After DNA extraction, tcdA, tcdB and binary toxin (CDTa/CDTb) genes were detected by PCR, and PCR-ribotyping performed. From a total of 913 stool samples collected during 2013–2014, 775 were included in the study. The frequency of A/B toxins-positive samples was 9.7% (75/775). A total of 143 isolates of C.difficile were recovered from culture, 110 (76.9%) produced cytotoxic effect in cell culture, 100 (69.9%) were tcdA+/tcdB+, 11 (7.7%) tcdA-/tcdB+, 32 (22.4%) tcdA-/tcdB- and 25 (17.5%) CDTa+/CDTb+. From 37 ribotypes identified, ribotypes 591 (20%), 106 (9%) and 002 (7.9%) were the most prevalent; only one isolate corresponded to ribotype 027, four to ribotype 078 and four were new ribotypes (794,795, 804,805). All isolates were susceptible to vancomycin and metronidazole, while 85% and 7.7% were resistant to clindamycin and moxifloxacin, respectively. By multivariate analysis, significant risk factors associated to CDI were, staying in orthopedic service, exposure to third-generation cephalosporins and staying in an ICU before CDI symptoms; moreover, steroids showed to be a protector factor. These results revealed new C. difficile ribotypes and a high diversity profile circulating in Colombia different from those reported in America and European countries

Understanding the Potential Impact of Different Drug Properties On SARS-CoV-2 Transmission and Disease Burden: A Modelling Analysis

Background The public health impact of the COVID-19 pandemic has motivated a rapid search for potential therapeutics, with some key successes. However, the potential impact of different treatments, and consequently research and procurement priorities, have not been clear. Methods Using a mathematical model of SARS-CoV-2 transmission, COVID-19 disease and clinical care, we explore the public-health impact of different potential therapeutics, under a range of scenarios varying healthcare capacity, epidemic trajectories; and drug efficacy in the absence of supportive care. Results The impact of drugs like dexamethasone (delivered to the most critically-ill in hospital and whose therapeutic benefit is expected to depend on the availability of supportive care such as oxygen and mechanical ventilation) is likely to be limited in settings where healthcare capacity is lowest or where uncontrolled epidemics result in hospitals being overwhelmed. As such, it may avert 22% of deaths in high-income countries but only 8% in low-income countries (assuming R=1.35). Therapeutics for different patient populations (those not in hospital, early in the course of infection) and types of benefit (reducing disease severity or infectiousness, preventing hospitalisation) could have much greater benefits, particularly in resource-poor settings facing large epidemics. Conclusions Advances in the treatment of COVID-19 to date have been focussed on hospitalised-patients and predicated on an assumption of adequate access to supportive care. Therapeutics delivered earlier in the course of infection that reduce the need for healthcare or reduce infectiousness could have significant impact, and research into their efficacy and means of delivery should be a priority