Development of Genetic Tools for the Manipulation of the Planctomycetes

Bacteria belonging to the Planctomycetes, Verrucomicrobia, Chlamydiae (PVC) superphylum are of interest for biotechnology, evolutionary cell biology, ecology, and human health. Some PVC species lack a number of typical bacterial features while others possess characteristics that are usually more associated to eukaryotes or archaea. For example, the Planctomycetes phylum is atypical for the absence of the FtsZ protein and for the presence of a developed endomembrane system. Studies of the cellular and molecular biology of these infrequent characteristics are currently limited due to the lack of genetic tools for most of the species. So far, genetic manipulation in Planctomycetes has been described in Planctopirus limnophila only. Here, we show a simple approach that allows mutagenesis by homologous recombination in three different planctomycetes species (i.e., Gemmata obscuriglobus, Gimesia maris, and Blastopirellula marina), in addition to P. limnophila, thus extending the repertoire of genetically modifiable organisms in this superphylum. Although the Planctomycetes show high resistance to most antibiotics, we have used kanamycin resistance genes in G. obscuriglobus, P. limnophila, and G. maris, and tetracycline resistance genes in B. marina, as markers for mutant selection. In all cases, plasmids were introduced in the strains by mating or electroporation, and the genetic modification was verified by Southern Blotting analysis. In addition, we show that the green fluorescent protein (gfp) is expressed in all four backgrounds from an Escherichia coli promoter. The genetic manipulation achievement in four phylogenetically diverse planctomycetes will enable molecular studies in these strains, and opens the door to developing genetic approaches not only in other planctomycetes but also other species of the superphylum, such as the Lentisphaerae.ER-M and DPD are supported by the Spanish Ministry of Economy and Competitivity (Grant BFU2013-40866-P) and the Junta de Andalucía (CEIC Grant C2A program to DPD). IC and ES are supported by the Spanish Ministry of Economy and Competitivity (Grant BIO2014-57545-R).Peer reviewedPeer Reviewe

Neuropsiquiatría clínica de la epilepsia: La amnesia epiléptica transitoria. A propósito de un caso.

Introducción: Muchos pacientes epilépticos refieren problemas mnésicos. Encontramos un grupo de pacientes que presentan episodios amnésicos de repetición y problemas en la memoria autobiográfica. Este cuadro clínico ha sido definido como “amnesia epiléptica transitoria”. Tras revisar la literatura, describimos las características de esta entidad nosológica con detalle.Caso clínico: Informamos de un caso de amnesia epiléptica transitoria en un varón de 58 años, con un diagnóstico previo de epilepsia y cuadro afectivo, que presenta episodios recurrentes y transitorios de amnesia mostrando el resto de sus funciones cognitivas preservadas. Durante los ataques se describen automatismos oro-alimentarios, breves ataques de estupor sin pérdida de consciencia ni desconexión con el medio.Conclusiones: La amnesia epiléptica transitoria es un subtipo de epilepsia del lóbulo temporal bien definido pero infradiagnosticado. En casos de pacientes que refieren  únicamente  problemas  de  memoria,  sobre todo si son episodios de amnesia breves y recurrentes, y en aquéllos pacientes que refieran deficiencias en la memoria remota, la amnesia epiléptica transitoria debe ser considerada como una opción diagnóstica seria. El tratamiento con fármacos anticomiciales produce muy buenos resultados

Neuropsiquiatría clínica de la epilepsia: La amnesia epiléptica transitoria. A propósito de un caso.

Introducción: Muchos pacientes epilépticos refieren problemas mnésicos. Encontramos un grupo de pacientes que presentan episodios amnésicos de repetición y problemas en la memoria autobiográfica. Este cuadro clínico ha sido definido como “amnesia epiléptica transitoria”. Tras revisar la literatura, describimos las características de esta entidad nosológica con detalle.Caso clínico: Informamos de un caso de amnesia epiléptica transitoria en un varón de 58 años, con un diagnóstico previo de epilepsia y cuadro afectivo, que presenta episodios recurrentes y transitorios de amnesia mostrando el resto de sus funciones cognitivas preservadas. Durante los ataques se describen automatismos oro-alimentarios, breves ataques de estupor sin pérdida de consciencia ni desconexión con el medio.Conclusiones: La amnesia epiléptica transitoria es un subtipo de epilepsia del lóbulo temporal bien definido pero infradiagnosticado. En casos de pacientes que refieren  únicamente  problemas  de  memoria,  sobre todo si son episodios de amnesia breves y recurrentes, y en aquéllos pacientes que refieran deficiencias en la memoria remota, la amnesia epiléptica transitoria debe ser considerada como una opción diagnóstica seria. El tratamiento con fármacos anticomiciales produce muy buenos resultados

Trastorno obsesivo compulsivo con tics motores y verbales, trastorno de acumulación y síndrome del acento extranjero sin afasia: comunicación de un caso y revisión bibliográfica

Informamos sobre un caso de trastorno obsesivo compulsivo (TOC), con tics motores y verbales, conducta acumuladora y síndrome del acento extranjero (dialecto regional) sin afasia, en un nativo español diestro que presentó una hemorragia en ganglios basales izquierdos. Su TOC y conducta acumuladora, que precedieron a la hemorragia cerebral, se diagnosticaron a raíz de un ingreso hospitalario, ya que previamente no había estado en contacto con ningún servicio psiquiátrico. Una lesión limitada al núcleo estriado izquierdo le provocó una variante regional del síndrome del acento extranjero (SAE). El paciente respondió adecuadamente al tratamiento. En nuestro conocimiento, éste es el primer caso de SAE en un hispanoparlante y también en un individuo que presentaba previamente TOC, tics y conducta acumuladora. Revisamos la literatura científica de estos trastorno

The H-ATOMIC Criteria for the Etiologic Classification of Patients with Intracerebral Hemorrhage

Background and Purpose There are no generally accepted criteria for the etiologic classification of intracerebral hemorrhage (ICH). For this reason, we have developed a set of etiologic criteria and have applied them to a large number of patients to determine their utility. Methods The H-ATOMIC classification includes 7 etiologic categories: Hypertension, cerebral Amyloid angiopathy, Tumour, Oral anticoagulants, vascular Malformation, Infrequent causes and Cryptogenic. For each category, the etiology is scored with three degrees of certainty: Possible(3), Probable(2) and Definite(1). Our aim was to perform a basic study consisting of neuroimaging, blood tests, and CT-angio when a numerical score (SICH) suggested an underlying structural abnormality. Combinations of >1 etiologic category for an individual patient were acceptable. The criteria were evaluated in a multicenter and prospective study of consecutive patients with spontaneous ICH. Results Our study included 439 patients (age 70.8 ± 14.5 years; 61.3% were men). A definite etiology was achieved in 176 (40.1% of the patients: Hypertension 28.2%, cerebral Amyloid angiopathy 0.2%, Tumour 0.2%, Oral anticoagulants 2.2%, vascular Malformation 4.5%, Infrequent causes 4.5%). A total of 7 patients (1.6%) were cryptogenic. In the remaining 58.3% of the patients, ICH was attributable to a single (n = 56, 12.7%) or the combination of 2 (n = 200, 45.5%) possible/probable etiologies. The most frequent combinations of etiologies involved possible hypertension with possible CAA (H3A3, n = 38) or with probable CAA (H3A2, n = 29), and probable hypertension with probable OA (H2O2, n = 27). The most frequent category with any degree of certainty was hypertension (H1+2+3 = 80.6%) followed by cerebral amyloid angiopathy (A1+2+3 = 30.9%). Conclusions According to our etiologic criteria, only about 40% patients received a definite diagnosis, while in the remaining patients ICH was attributable to a single possible/probable etiology or to more than one possible/probable etiology. The use of these criteria would likely help in the management of patients with ICH.This work was supported by Ministery of Health-Instituto de Salud Carlos III: RETICS (Redes temáticas de Investigación Cooperativa) INVICTUS RD012/0014 (JM-F, PC-R, AM-D, LP-S, RD-M), FEDER (Fondo Europeo de Desarrollo Regional)

Detection of kinase domain mutations in BCR::ABL1 leukemia by ultra-deep sequencing of genomic DNA

The screening of the BCR::ABL1 kinase domain (KD) mutation has become a routine analysis in case of warning/failure for chronic myeloid leukemia (CML) and B-cell precursor acute lymphoblastic leukemia (ALL) Philadelphia (Ph)-positive patients. In this study, we present a novel DNA-based next-generation sequencing (NGS) methodology for KD ABL1 mutation detection and monitoring with a 1.0E−4 sensitivity. This approach was validated with a well-stablished RNA-based nested NGS method. The correlation of both techniques for the quantification of ABL1 mutations was high (Pearson r = 0.858, p < 0.001), offering DNA-DeepNGS a sensitivity of 92% and specificity of 82%. The clinical impact was studied in a cohort of 129 patients (n = 67 for CML and n = 62 for B-ALL patients). A total of 162 samples (n = 86 CML and n = 76 B-ALL) were studied. Of them, 27 out of 86 harbored mutations (6 in warning and 21 in failure) for CML, and 13 out of 76 (2 diagnostic and 11 relapse samples) did in B-ALL patients. In addition, in four cases were detected mutation despite BCR::ABL1 < 1%. In conclusion, we were able to detect KD ABL1 mutations with a 1.0E−4 sensitivity by NGS using DNA as starting material even in patients with low levels of disease.Tis project was funded in part by CRIS CANCER FOUNDATION

Detection of kinase domain mutations in BCR::ABL1 leukemia by ultra-deep sequencing of genomic DNA

The screening of the BCR::ABL1 kinase domain (KD) mutation has become a routine analysis in case of warning/failure for chronic myeloid leukemia (CML) and B-cell precursor acute lymphoblastic leukemia (ALL) Philadelphia (Ph)-positive patients. In this study, we present a novel DNA-based next-generation sequencing (NGS) methodology for KD ABL1 mutation detection and monitoring with a 1.0E-4 sensitivity. This approach was validated with a well-stablished RNA-based nested NGS method. The correlation of both techniques for the quantification of ABL1 mutations was high (Pearson r = 0.858, p < 0.001), offering DNA-DeepNGS a sensitivity of 92% and specificity of 82%. The clinical impact was studied in a cohort of 129 patients (n = 67 for CML and n = 62 for B-ALL patients). A total of 162 samples (n = 86 CML and n = 76 B-ALL) were studied. Of them, 27 out of 86 harbored mutations (6 in warning and 21 in failure) for CML, and 13 out of 76 (2 diagnostic and 11 relapse samples) did in B-ALL patients. In addition, in four cases were detected mutation despite BCR::ABL1 < 1%. In conclusion, we were able to detect KD ABL1 mutations with a 1.0E-4 sensitivity by NGS using DNA as starting material even in patients with low levels of disease

COVID-19 Severity and Survival over Time in Patients with Hematologic Malignancies: A Population-Based Registry Study

Mortality rates for COVID-19 have declined over time in the general population, but data in patients with hematologic malignancies are contradictory. We identified independent prognostic factors for COVID-19 severity and survival in unvaccinated patients with hematologic malignancies, compared mortality rates over time and versus non-cancer inpatients, and investigated post COVID-19 condition. Data were analyzed from 1166 consecutive, eligible patients with hematologic malignancies from the population-based HEMATO-MADRID registry, Spain, with COVID-19 prior to vaccination roll-out, stratified into early (February–June 2020; n = 769 (66%)) and later (July 2020–February 2021; n = 397 (34%)) cohorts. Propensity-score matched non-cancer patients were identified from the SEMI-COVID registry. A lower proportion of patients were hospitalized in the later waves (54.2%) compared to the earlier (88.6%), OR 0.15, 95%CI 0.11–0.20. The proportion of hospitalized patients admitted to the ICU was higher in the later cohort (103/215, 47.9%) compared with the early cohort (170/681, 25.0%, 2.77; 2.01–3.82). The reduced 30-day mortality between early and later cohorts of non-cancer inpatients (29.6% vs. 12.6%, OR 0.34; 0.22–0.53) was not paralleled in inpatients with hematologic malignancies (32.3% vs. 34.8%, OR 1.12; 0.81–1.5). Among evaluable patients, 27.3% had post COVID-19 condition. These findings will help inform evidence-based preventive and therapeutic strategies for patients with hematologic malignancies and COVID-19 diagnosis.Depto. de MedicinaFac. de MedicinaTRUEFundación Madrileña de Hematología y HemoterapiaFundación Leucemia y LinfomaAsociación Madrileña de Hematología y Hemoterapiapu

Desafíos actuales de la educación superior

El presente libro corresponde al trabajo investigativo realizado por sus autores frente a los desafíos que enfrenta y enfrentará la educación en tiempos actuales (provisional)