Incidencia de infección por SARS-CoV-2 en contactos estrechos. Seguimiento telefónico en Atención Primaria / Incidence of SARS-CoV-2 infection in close contacs. Telephone follow-up in Primary Care

Objetivo: Estudiar mediante un protocolo de seguimiento telefónico la incidencia de infección en los contactos estrechos de pacientes con infección por SARS-CoV-2 tras la recomendación de aislamiento y cuarentena en el domicilio.Pacientes y métodosEstudio de cohortes, con 124 pacientes mayores de 15 años, incluidos consecutivamente, asintomáticos en el momento de la visita, con contacto estrecho (<2 metros) con casos confirmados o posibles de infección por SARS-CoV-2. La intervención consistió en aislamiento y cuarentena en domicilio durante 2 semanas, contactando telefónicamente los días 2, 4, 7 y 14 tras el contacto. El evento de interés fue la aparición de clínica compatible con infección por SARS-CoV-2.ResultadosLa edad media fue de 45,1 años (55,6% mujeres). Se realizaron 328 llamadas telefónicas (media de 2,6 llamadas por paciente). Tras 2 semanas de seguimiento desarrollaron síntomas 6 pacientes, confirmados serológicamente o por PCR en 3 de ellos (2,4%; IC 95%: 0,8-6,9).ConclusionesLa incidencia de infección por SARS-CoV-2 en los contactos estrechos es baja a las 2 semanas tras el aislamiento y cuarentena domiciliarios. (AU)

Objective: To study the incidence of infection in close contacts with patiens with SARS-CoV-2 infection using a telephone monitoring protocol after the recommendation of isolation and quarantine at home.Patients and methodsCohort study, with 124 patients older than 15 years, included consecutively, asymptomatic at the time of the consultation, who had had close contact (<2 meters) with confirmed or possible cases of SARS-CoV-2 infection. The intervention consisted of 2 weeks of home isolation and quarantine, contacting by phone on days, 2, 4, 7 and 14 after the contact. The event of interest was the appereance of symptoms compatible with SARS-CoV-2 infection.ResultsThe average age was 45.1 years (55.6% women); 328 phone calls were made (average 2.6 calls for patient). After 2 weeks of follow-up, 6 patients developed symptoms, confirming serologically or by PCR in 3 of them (2.4%, CI 95%: 0.8-6.9).ConclusionsThe incidence of SARS-CoV-2 infection in close contacts is low 2 weeks after home isolation and quarantine at home. (AU)

Incidence of SARS-CoV-2 infection in close contacs. Telephone follow-up in Primary Care.

OBJECTIVE: To study the incidence of infection in close contacts with patiens with SARS-CoV-2 infection using a telephone monitoring protocol after the recommendation of isolation and quarantine at home. PATIENTS AND METHODS: Cohort study, with 124 patients older than 15 years, included consecutively, asymptomatic at the time of the consultation, who had had close contact (<2 m) with confirmed or possible cases of SARS-CoV-2 infection. The intervention consisted of 2 weeks of home isolation and quarantine, contacting by phone on days, 2, 4, 7 and 14 after the contact. The event of interest was the appereance of symptoms compatible with SARS-CoV-2 infection. RESULTS: The average age was 45.1 years (55.6% women); 328 phone calls were made (average 2.6 calls for patient). After 2 weeks of follow-up, 6 patients developed symptoms, confirming serologically or by PCR in 3 of them (2.4%, CI 95%: 0.8-6.9). CONCLUSIONS: The incidence of SARS-CoV-2 infection in close contacts is low 2 weeks after home isolation and quarantine at home.

OBJETIVO: Estudiar mediante un protocolo de seguimiento telefónico la incidencia de infección en los contactos estrechos de pacientes con infección por SARS-CoV-2 tras la recomendación de aislamiento y cuarentena en el domicilio. PACIENTES Y MÉTODOS: Estudio de cohortes, con 124 pacientes mayores de 15 años, incluidos consecutivamente, asintomáticos en el momento de la visita, con contacto estrecho (<2 metros) con casos confirmados o posibles de infección por SARS-CoV-2. La intervención consistió en aislamiento y cuarentena en domicilio durante 2 semanas, contactando telefónicamente los días 2, 4, 7 y 14 tras el contacto. El evento de interés fue la aparición de clínica compatible con infección por SARS-CoV-2. RESULTADOS: La edad media fue de 45,1 años (55,6% mujeres). Se realizaron 328 llamadas telefónicas (media de 2,6 llamadas por paciente). Tras 2 semanas de seguimiento desarrollaron síntomas 6 pacientes, confirmados serológicamente o por PCR en 3 de ellos (2,4%; IC 95%: 0,8­6,9). CONCLUSIONES: La incidencia de infección por SARS-CoV-2 en los contactos estrechos es baja a las 2 semanas tras el aislamiento y cuarentena domiciliarios.

[Incidence of SARS-CoV-2 infection in close contacs. Telephone follow-up in Primary Care]. / Incidencia de infección por SARS-CoV-2 en contactos estrechos. Seguimiento telefónico en Atención Primaria.

OBJECTIVE: To study the incidence of infection in close contacts with patiens with SARS-CoV-2 infection using a telephone monitoring protocol after the recommendation of isolation and quarantine at home. PATIENTS AND METHODS: Cohort study, with 124 patients older than 15 years, included consecutively, asymptomatic at the time of the consultation, who had had close contact (<2 meters) with confirmed or possible cases of SARS-CoV-2 infection. The intervention consisted of 2 weeks of home isolation and quarantine, contacting by phone on days, 2, 4, 7 and 14 after the contact. The event of interest was the appereance of symptoms compatible with SARS-CoV-2 infection. RESULTS: The average age was 45.1 years (55.6% women); 328 phone calls were made (average 2.6 calls for patient). After 2 weeks of follow-up, 6 patients developed symptoms, confirming serologically or by PCR in 3 of them (2.4%, CI 95%: 0.8-6.9). CONCLUSIONS: The incidence of SARS-CoV-2 infection in close contacts is low 2 weeks after home isolation and quarantine at home.

Successful engraftment of gene-corrected hematopoietic stem cells in non-conditioned patients with Fanconi anemia.

Fanconi anemia (FA) is a DNA repair syndrome generated by mutations in any of the 22 FA genes discovered to date1,2. Mutations in FANCA account for more than 60% of FA cases worldwide3,4. Clinically, FA is associated with congenital abnormalities and cancer predisposition. However, bone marrow failure is the primary pathological feature of FA that becomes evident in 70-80% of patients with FA during the first decade of life5,6. In this clinical study (ClinicalTrials.gov, NCT03157804 ; European Clinical Trials Database, 2011-006100-12), we demonstrate that lentiviral-mediated hematopoietic gene therapy reproducibly confers engraftment and proliferation advantages of gene-corrected hematopoietic stem cells (HSCs) in non-conditioned patients with FA subtype A. Insertion-site analyses revealed the multipotent nature of corrected HSCs and showed that the repopulation advantage of these cells was not due to genotoxic integrations of the therapeutic provirus. Phenotypic correction of blood and bone marrow cells was shown by the acquired resistance of hematopoietic progenitors and T lymphocytes to DNA cross-linking agents. Additionally, an arrest of bone marrow failure progression was observed in patients with the highest levels of gene marking. The progressive engraftment of corrected HSCs in non-conditioned patients with FA supports that gene therapy should constitute an innovative low-toxicity therapeutic option for this life-threatening disorder.

Generation of a High Number of Healthy Erythroid Cells from Gene-Edited Pyruvate Kinase Deficiency Patient-Specific Induced Pluripotent Stem Cells.

Pyruvate kinase deficiency (PKD) is a rare erythroid metabolic disease caused by mutations in the PKLR gene. Erythrocytes from PKD patients show an energetic imbalance causing chronic non-spherocytic hemolytic anemia, as pyruvate kinase defects impair ATP production in erythrocytes. We generated PKD induced pluripotent stem cells (PKDiPSCs) from peripheral blood mononuclear cells (PB-MNCs) of PKD patients by non-integrative Sendai viral vectors. PKDiPSCs were gene edited to integrate a partial codon-optimized R-type pyruvate kinase cDNA in the second intron of the PKLR gene by TALEN-mediated homologous recombination (HR). Notably, we found allele specificity of HR led by the presence of a single-nucleotide polymorphism. High numbers of erythroid cells derived from gene-edited PKDiPSCs showed correction of the energetic imbalance, providing an approach to correct metabolic erythroid diseases and demonstrating the practicality of this approach to generate the large cell numbers required for comprehensive biochemical and metabolic erythroid analyses.

Development of an in vitro model for the simultaneous study of the efficacy and hematotoxicity of antileukemic compounds.

Hematopoietic system displays a wide spectrum of cell populations hierarchically organized in the bone marrow. Homeostasis in this system requires equilibrium between the self-renewal of the stem cells and their capacity of differentiation. Any failure on this equilibrium could lead to fatal consequences, such as the development of leukemia. Due to its rapid rate of renewal, hematopoietic tissue is a major target for antitumoral compounds and often becomes a dose limiting factor in the development of antineoplastics. Our aim was to develop an in vitro model for predicting the efficacy of antitumoral compounds on leukemic cells and their toxic effects on the healthy hematopoietic cells. The mouse myelomonocytic leukemia WEHI-3b was transduced with a lentiviral vector for expressing the green fluorescence protein. Mixed semisolid clonogenic cultures of transduced WEHI-3b and murine bone marrow cells were exposed to five pharmaceuticals: daunorubicin (positive control), atropine sulphate (negative control) and three in different stages of clinical development (trabectedin, Zalypsis(®) and PM01183). Colonies of leukemic cells were distinguishable from healthy CFU-GM under fluorescence microscope. The sensitivity of leukemic cells to daunorubicin, trabectedin, Zalypsis(®) and PM01183 was higher compared to healthy cells. The effect of a non-antitumoral compound, atropine sulphate, was the same on both populations. Our results show that this in vitro model is a valuable tool for studying the effect of antitumoral compounds in both tumoral and normal hematopoietic cells under the same toxic microenvironment and could safe time and facilitate the reduction of the number of animals used in preclinical development of pharmaceuticals.

Prevalidation of the rat CFU-GM assay for in vitro toxicology applications.

In vitro haematotoxicity assays are thought to have the potential to significantly reduce and refine the use of animals for haematotoxicity testing. These assays are used successfully in all types of studies - however, their use is not so common in human toxicology studies in the preclinical setting, as they are not required for regulatory testing in this case. Furthermore, these assays could play a key role in bridging the gap between preclinical toxicology studies in animal models and clinical investigations. In previous studies, the Colony Forming Unit-Granulocyte Macrophage (CFU-GM) assay has been validated for testing drug haematotoxicity (with both mouse bone-marrow and human cord blood) and for predicting the in vivo human maximal tolerated dose (MTD) by adjusting in vivo data on mouse toxicity. Recently, a Colony Forming Unit-Megakaryocyte (CFU-MK) assay has also been prevalidated for testing drug toxicity toward megakaryocytes. The rat CFU-GM assay has been used by many researchers for its ability to evaluate in vitro haematotoxicity. Although it is not yet available, a standardised procedure for data comparison could be very important, since the rat is the most widely-used species for the in vivo testing of toxicants. This report presents the results of the prevalidation study developed to analyse the intra-laboratory and inter-laboratory variability of a standardised operating procedure for this assay and its performance for the in vitro determination of the inhibitory concentration (IC) values of drugs on rat myeloid progenitors (CFU-GM). The results demonstrate that the CFU-GM assay can be performed with cryopreserved rat bone-marrow cells (rBMC). The assay represents a useful tool for evaluating the toxicity of a compound, in terms of both relative toxicity (when different molecules are compared) and the prediction of the degree of in vivo toxicity. The use of this assay could greatly reduce the number of rats used in experimental procedures, and could also contribute to the accumulation of more toxicity data on compounds to be registered according to the criteria established by the European Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) programme.

In vitro sensitivity of granulo-monocytic progenitors as a new toxicological cell system and endpoint in the ACuteTox Project.

The ACuteTox Project (part of the EU 6th Framework Programme) was started up in January 2005. The aim of this project is to develop a simple and robust in vitro strategy for prediction of human acute systemic toxicity, which could replace animal tests used for regulatory purposes. Our group is responsible for the characterization of the effect of the reference chemicals on the hematopoietic tissue. CFU-GM assay based on the culture of human mononuclear cord blood cells has been used to characterize the effects of the selected compounds on the myeloid progenitors. Previous results have shown the relevance of the CFU-GM assay for the prediction of human acute neutropenia after treatment of antitumoral compounds, and this assay has been recently approved by the ECVAM's Scientific Advisory Committee. Among the compounds included in the study there were pharmaceuticals, environmental pollutants and industrial chemicals. Eleven out of 55 chemicals did not show any cytotoxic effect at the maximum concentration tested. The correlation coefficients of CFU-GM IC50, IC70 and IC90 values with human LC50 values (50% lethal concentration calculated from time-related sublethal and lethal human blood concentrations) were 0.4965, 0.5106 and 0.5142 respectively. Although this correlation is not improve respect to classical in vitro basal cytotoxicity tests such as 3T3 Neutral Red Uptake, chemicals which deviate substantially in the correlation with these assays (colchicine, digoxin, 5-Fluorouracil and thallium sulfate) fitted very well in the linear regression analysis of the CFU-GM progenitors. The results shown in the present study indicate that the sensitivity of CFU-GM progenitors correlates better than the sensitivity of HL-60 cells with human LC50 values and could help to refine the predictability for human acute systemic toxicity when a given chemical may affect to the hematopoietic myeloid system.

Impact of the clinical context on the 14-3-3 test for the diagnosis of sporadic CJD.

BACKGROUND: The 14-3-3 test appears to be a valuable aid for the clinical diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) in selected populations. However, its usefulness in routine practice has been challenged. In this study, the influence of the clinical context on the performance of the 14-3-3 test for the diagnosis of sCJD is investigated through the analysis of a large prospective clinical series. METHODS: Six hundred seventy-two Spanish patients with clinically suspected sCJD were analyzed. Clinical classification at sample reception according to the World Health Organization's (WHO) criteria (excluding the 14-3-3 test result) was used to explore the influence of the clinical context on the pre-test probabilities, and positive (PPV) and negative (NPV) predictive values of the 14-3-3 test. RESULTS: Predictive values of the test varied greatly according to the initial clinical classification: PPV of 98.8%, 96.5% and 45.0%, and NPV of 26.1%, 66.6% and 100% for probable sCJDi (n = 115), possible sCJDi (n = 73) and non-sCJDi (n = 484) cases, respectively. According to multivariate and Bayesian analyses, these values represent an improvement of diagnostic certainty compared to clinical data alone. CONCLUSION: In three different contexts of sCJD suspicion, the 14-3-3 assay provides useful information complementary to clinical and electroencephalographic (EEG) data. The test is most useful supporting a clinical impression, whilst it may show deceptive when it is not in agreement with clinical data.