5′ flanking region of var genes nucleate histone modification patterns linked to phenotypic inheritance of virulence traits in malaria parasites

In the human malaria parasite Plasmodium falciparum antigenic variation facilitates long-term chronic infection of the host. This is achieved by sequential expression of a single member of the 60-member var family. Here we show that the 5′ flanking region nucleates epigenetic events strongly linked to the maintenance of mono-allelic var gene expression pattern during parasite proliferation. Tri- and dimethylation of histone H3 lysine 4 peak in the 5′ upstream region of transcribed var and during the poised state (non-transcribed phase of var genes during the 48 h asexual life cycle), ‘bookmarking’ this member for re-activation at the onset of the next cycle. Histone H3 lysine 9 trimethylation acts as an antagonist to lysine 4 methylation to establish stably silent var gene states along the 5′ flanking and coding region. Furthermore, we show that competition exists between H3K9 methylation and H3K9 acetylation in the 5′ flanking region and that these marks contribute epigenetically to repressing or activating var gene expression. Our work points to a pivotal role of the histone methyl mark writing and reading machinery in the phenotypic inheritance of virulence traits in the malaria parasite

Chronic lymphocytic leukemia patients with IGH translocations are characterized by a distinct genetic landscape with prognostic implications

Chromosome 14q32 rearrangements/translocations involving the immunoglobulin heavy chain (IGH) are rarely detected in chronic lymphocytic leukemia (CLL). The prognostic significance of the IGH translocation is controversial and its mutational profile remains unknown. Here, we present for the first time a comprehensive next-generation sequencing (NGS) analysis of 46 CLL patients with IGH rearrangement (IGHR-CLLs) and we demonstrate that IGHR-CLLs have a distinct mutational profile with recurrent mutations in NOTCH1, IGLL5, POT1, BCL2, FBXW7, ZMYM3, MGA, BRAF and HIST1H1E genes. Interestingly, BCL2 and FBXW7 mutations were significantly associated with this subgroup and almost half of BCL2, IGLL5 and HISTH1E mutations reported were previously identified in non-Hodgkin lymphomas. Notably, IGH/BCL2 rearrangements were associated with a lower mutation frequency and carried BCL2 and IGLL5 mutations, while the other IGHR-CLLs had mutations in genes related to poor prognosis (NOTCH1, SF3B1 and TP53) and shorter time to first treatment (TFT). Moreover, IGHR-CLLs patients showed a shorter TFT than CLL patients carrying 13q-, normal fluorescence in situ hybridization (FISH) and +12 CLL, being this prognosis particularly poor when NOTCH1, SF3B1, TP53, BIRC3 and BRAF were also mutated. The presence of these mutations not only was an independent risk factor within IGHR-CLLs, but also refined the prognosis of low-risk cytogenetic patients (13q-/normal FISH). Hence, our study demonstrates that IGHR-CLLs have a distinct mutational profile from the majority of CLLs and highlights the relevance of incorporating NGS and the status of IGH by FISH analysis to refine the risk-stratification CLL model

Treatment patterns and outcomes among nontransplant newly diagnosed multiple myeloma patients in Spain

Aim: To describe treatment patterns and outcomes in nontransplant newly diagnosed multiple myeloma (NDMM) patients in Spain. Methods: This retrospective study included two cohorts of NDMM patients diagnosed between 1 January 2012 to 31 December 2013 and 1 April 2016 to 31 March 2017. Results: Among 113 patients, proteasome inhibitor (PI) + alkylator combinations (49%) and PI-based regimens without an alkylator (30%) were the most common first-line (1L) therapies. Use of PI + immunomodulatory drug-based regimens increased between the cohorts; PI-based regimens without an alkylator/immunomodulatory drug decreased. Use of 1L oral regimens was low but increased over time; use of maintenance therapy was low across both periods. Median 1L duration of treatment was 6.9 months. Conclusion: Short 1L duration of treatment and low use of 1L oral regimens and maintenance therapy highlight unmet needs in NDMM. © 2021 The Authors

ACTIVIDAD AMEBICIDA, ANTIOXIDANTE Y PERFIL FITOQUÍMICO DE EXTRACTOS METANÓLICOS DE ASTROPHYTUM MYRIOSTIGMA OBTENIDOS DE CULTIVO DE CALLO Y DEL CACTUS SILVESTRE

Methanolic extracts were obtained fromboth in vitro and ex vitro tissues of Astrophytum myriostigma to develop a phytochemical profile, probe antioxidant activity(DPPH 2, 2-diphenyl-1-picryl-hydrazyl)and evaluate amoebicid activity againstEntamoeba histolytica HM1-IMSS. Thephytochemical profile results indicate thepresence of different secondary metabolitesin callus and stem; whereas the callus hasphenolic oxidrils and flavonoids, these arenot present in the stem, which may causethe difference in the antioxidant activitybetween callus with an IC50 of 199.19μg/mL and stem with 3961.39 μg/mL. Theamoebicid activity was satisfactory withan IC50 of <100μg/mL for both extracts.The presence of these metabolites in callusof Astrophytum myriostigma is probablydue to conditions and nutrients present inmedia.Se determinó la actividad amebicida, antioxidante y perfil fitoquímico de extractosmetanólicos de callo cultivado in vitro ysecciones de tallo de planta ex vitro deAstrophytum myriostigma. Se obtuvieronextractos metanólicos de ambos tejidos paraevaluar la actividad amebicida sobre Entamoeba histolytica HM1-IMSS, la actividadantioxidante (DPPH 2,2-diphenyl-1-picrylhydrazyl) y su perfil fitoquímico. Paraambos extractos, la actividad amebicidaresultó satisfactoria con una concentracióninhibitoria media menor de 100 μg/mL. Losresultados del perfil fitoquímico indican lapresencia de metabolitos secundarios diferentes en callo y tallo, mientras que el callo,posee oxidrilos fenólicos y flavonoides,en tallo no están presentes, lo que puedeocasionar la diferencia en los resultadosde la actividad antioxidante, para callo unaconcentración efectiva media de 199.19μg/mL y para tallo de 3961.39 μg/mL. Lapresencia de estos metabolitos en callo deAstrophytum myriostigma, se debe probablemente a las condiciones y a los nutrientespresentes en el medio in vitro

Inadequate use of antibiotics in the covid-19 era: effectiveness of antibiotic therapy

Background: Since December 2019, the COVID-19 pandemic has changed the concept of medicine. This work aims to analyze the use of antibiotics in patients admitted to the hospital due to SARS-CoV-2 infection. Methods: This work analyzes the use and effectiveness of antibiotics in hospitalized patients with COVID-19 based on data from the SEMI-COVID-19 registry, an initiative to generate knowledge about this disease using data from electronic medical records. Our primary endpoint was all-cause in-hospital mortality according to antibiotic use. The secondary endpoint was the effect of macrolides on mortality. Results: Of 13, 932 patients, antibiotics were used in 12, 238. The overall death rate was 20.7% and higher among those taking antibiotics (87.8%). Higher mortality was observed with use of all antibiotics (OR 1.40, 95% CI 1.21–1.62; p <.001) except macrolides, which had a higher survival rate (OR 0.70, 95% CI 0.64–0.76; p <.001). The decision to start antibiotics was influenced by presence of increased inflammatory markers and any kind of infiltrate on an x-ray. Patients receiving antibiotics required respiratory support and were transferred to intensive care units more often. Conclusions: Bacterial co-infection was uncommon among COVID-19 patients, yet use of antibiotics was high. There is insufficient evidence to support widespread use of empiric antibiotics in these patients. Most may not require empiric treatment and if they do, there is promising evidence regarding azithromycin as a potential COVID-19 treatment. © 2021, The Author(s)

Sobre la presencia de Teucrium pumilum y Teucrium libanitis (Lamiaceae) en la provincia de Valencia

Teucrium pumilum y T. libanitis son dos especies que han sido citadas como presentes en la provincia de Valencia desde mediados del siglo XX, para los yesos que afloran en la comarca del Valle de Ayora-Cofrentes. El análisis de las etiquetas de los pliegos de los herbarios no es concluyente; ningún pliego testigo certificaría la presencia de estas especies. La determinación de visu por parte de Rivas Goday, y su inclusión en una tabla de inventarios fitosociológica, son el único fundamento de su existencia en la zona en algún momento determinado de la reciente historia botánica española. Además, para T. pumilum, el pliego de herbario que tradicionalmente se ha asignado a la cita valenciana se trata de una confusión en la determinación por parte de algunos autores con su congénere T. carolipaui, siendo imposible al mismo tiempo asignar una localidad geográfica concreta a este material. Abstract. About the presence of Teucrium pumilum and Teucrium libanitis (Lamiaceae) in the Valencian province, Spain.- Teucrium pumilum and T. libanitis have been cited from Valencia province (Spain) since the middle of the XXth Century from the gypsic soils in the Valle de Ayora-Cofrentes shire. The analysis of specimens and labels is not conclusive; no specimen would certify the presence of any of these taxa. The field identification and their inclusion in a phyotosociological table (relevé) by Rivas Goday are the only basis of their presence in the territory in a particular moment of the recent Spanish botanical history. Additionally, the unique herbarium specimen from Valencia, impossible to assign a concrete geographic locality, which was traditionally assigned to T. pumilum by some authors, is actually its congeneric T. carolipaui. Resum. Sobre la presència de Teucrium pumilum i Teucrium libanitis (Lamiaceae) a la província de València.- Teucrium pumilum i T. libanitis són dues espècies que han estat citades com a presents a la província de València des de mitjans del segle XX, sobre els guixos que afloren a la comarca de la Vall d'Aiora-Cofrents. L'anàlisi de les etiquetes dels plecs dels herbaris no és concloent; cap plec testimoni certificaria la presència d'aquestes espècies. La determinació de visu per part de Rivas Goday, i la seva inclusió en una taula d'inventaris fitosociològica, són l'únic fonament de la seva existència a la zona en un moment determinat de la recent història botànica espanyola. A més, per a T. pumilum, el plec d'herbari que tradicionalment s' ha assignat a la cita valenciana es tracta d'una confusió en la determinació per part d'alguns autors amb el seu congènere T. carolipaui, essent impossible al mateix temps assignar una localitat geogràfica concreta a aquest material

BIOPROSPECCIÓN DE LA ACTIVIDAD ANTIMICÓTICA DE EXTRACTOS METANÓLICOS DE ARIOCARPUS KOTSCHOUBEYANUS Y ARIOCARPUS RETUSUS

The antifungal activity of methanolic extracts from Ariocarpus kotschoubeyanusand A. retusus, two native xerophytes ofnortheastern Mexico, was determined onthe fungal dermatophytes Trichophytontonsurans, Microsporum canis and Microsporum cookei. Putative active metabolites were identified by phytochemicalscreening using evidence for functionalgroups; determination of antifungal activity was conducted by the disk-diffusion platemethod as part of the process of understanding the chemistry and efficacy of theseplant products. Doses tested were 125,250 y 500 mg/mL. Extracts were positivefor carbonyl groups, phenolic oxhidrils,sterols and methyl sterols, coumarins, sesquiterpenlactones, saponins, flavonoids andalkaloids. Extract from A. retusus (stem)yielded the greatest activity against Trichophyton tonsurans and Microsporum cookei.Univariate analysis of variance revealedthat with regard to antifungal activity therewere significant differences between fungi,culture media and analyzed doses withrespect to positive control of ketoconazole.These results served to validate empiricalknowledge about these plants in traditionalmedicine. These studies form a basis forfurther research in the search for bioactivecompounds and the discovery of potentialherbal medicaments.Se determinó la actividad antifúngica invitro de extractos metanólicos de Ariocarpus kotschoubeyanus y Ariocarpusretusus, plantas xerófitas del noreste delpaís, usadas en la medicina tradicional,sobre los hongos dermatofitos Trichophytontonsurans, Microsporum canis y Microsporum cookei. Los metabolitos presentes seidentificaron por tamizaje fitoquímico empleando pruebas para grupos funcionales, ladeterminación de actividad antifúngica serealizó por el método de difusión en placaempleando microdiscos, con la finalidadde entender la química y eficacia de estosproductos vegetales, las dosis evaluadasde estos extractos fueron 125, 250 y 500mg/mL. Los extractos resultaron positivospara grupos carbonilo, oxidrilos fenólicos,esteroles y metilesteroles, cumarinas, sesquiterpenlactonas, saponinas, flavonoides yalcaloides. El extracto que presentó mayoractividad fue el de Ariocarpus retusus(tallo) contra Trichophyton tonsurans y Microsporum cookei. El análisis estadístico devarianza univariante reveló que en cuantoa la actividad antifúngica, existe diferencia significativa entre hongos, medios de cultivo y dosis evaluadas de los extractos conrespecto al control positivo de ketoconazol.Estos resultados sirvieron para validar elconocimiento empírico que se tiene acercade estas plantas en la medicina tradicional.Siendo estos estudios una base para nuevas investigaciones sobre la búsqueda decompuestos bioactivos y el descubrimientode medicamentos potenciales de productosherbales

Dissecting the role of TP53 alterations in del(11q) chronic lymphocytic leukemia

© 2021 The Authors.[Background]: Several genetic alterations have been identified as driver events in chronic lymphocytic leukemia (CLL) pathogenesis and oncogenic evolution. Concurrent driver alterations usually coexist within the same tumoral clone, but how the cooperation of multiple genomic abnormalities contributes to disease progression remains poorly understood. Specifically, the biological and clinical consequences of concurrent high-risk alterations such as del(11q)/ATM-mutations and del(17p)/TP53-mutations have not been established.[Methods]: We integrated next-generation sequencing (NGS) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 techniques to characterize the in vitro and in vivo effects of concurrent monoallelic or biallelic ATM and/or TP53 alterations in CLL prognosis, clonal evolution, and therapy response.[Results]: Targeted sequencing analysis of the co-occurrence of high-risk alterations in 271 CLLs revealed that biallelic inactivation of both ATM and TP53 was mutually exclusive, whereas monoallelic del(11q) and TP53 alterations significantly co-occurred in a subset of CLL patients with a highly adverse clinical outcome. We determined the biological effects of combined del(11q), ATM and/or TP53 mutations in CRISPR/Cas9-edited CLL cell lines. Our results showed that the combination of monoallelic del(11q) and TP53 mutations in CLL cells led to a clonal advantage in vitro and in in vivo clonal competition experiments, whereas CLL cells harboring biallelic ATM and TP53 loss failed to compete in in vivo xenotransplants. Furthermore, we demonstrated that CLL cell lines harboring del(11q) and TP53 mutations show only partial responses to B cell receptor signaling inhibitors, but may potentially benefit from ATR inhibition.[Conclusions]: Our work highlights that combined monoallelic del(11q) and TP53 alterations coordinately contribute to clonal advantage and shorter overall survival in CLL.Spanish Fondo de Investigaciones Sanitarias, Grant/Award Numbers: PI15/01471, PI18/01500); Fundación Memoria Don Samuel Solórzano Barruso, Grant/Award Number: RD12/0036/006

Clinical characteristics of patients hospitalized with COVID-19 in Spain: Results from the SEMI-COVID-19 Registry

Background: Spain has been one of the countries most affected by the COVID-19 pandemic. Objective: To create a registry of patients with COVID-19 hospitalized in Spain, in order to improve our knowledge of the clinical, diagnostic, therapeutic, and prognostic aspects of this disease. Methods: A multicentre retrospective cohort study, including consecutive patients hospitalized with confirmed COVID-19 throughout Spain. Epidemiological and clinical data, additional tests at admission and at seven days, treatments administered, and progress at 30 days of hospitalization were collected from electronic medical records. Results: Up to June 30th 2020, 15,111 patients from 150 hospitals were included. Their median age was 69.4 years (range: 18-102 years) and 57.2% were male. Prevalences of hypertension, dyslipidemia, and diabetes mellitus were 50.9%, 39.7%, and 19.4%, respectively. The most frequent symptoms were fever (84.2%) and cough (73.5%). High values of ferritin (73.5%), lactate dehydrogenase (73.9%), and D-dimer (63.8%), as well as lymphopenia (52.8%), were frequent. The most used antiviral drugs were hydroxychloroquine (85.6%) and lopinavir/ritonavir (61.4%); 33.1% developed respiratory distress. Overall mortality rate was 21.0%, with a marked increase with age (50-59 years: 4.7%, 60-69 years: 10.5%, 70-79 years: 26.9%, ≥ 80 years: 46.0%). Conclusions: The SEMI-COVID-19 Network provides data on the clinical characteristics of patients with COVID-19 hospitalized in Spain. Patients with COVID-19 hospitalized in Spain are mostly severe cases, as one in three patients developed respiratory distress and one in five patients died. These findings confirm a close relationship between advanced age and mortality.Antecedentes: España ha sido uno de los países más afectados por la pandemia de COVID-19.Objetivo: Crear un registro de pacientes hospitalizados en España por COVID-19 para mejorar nuestro conocimiento sobre los aspectos clínicos, diagnósticos, terapéuticos y pronósticos de esta enfermedad. Métodos: Estudio de cohorte retrospectiva, multicéntrico, que incluye pacientes consecutivos hospitalizados con COVID-19 confirmada en toda España. Se obtuvieron los datos epidemiológicos y clínicos, las pruebas complementarias al ingreso y a los 7 días de la admisión, los tratamientos administrados y la evolución a los 30 días de hospitalización de las historias clínicas electrónicas. Resultados: Hasta el 30 de junio de 2020 se incluyeron 15.111 pacientes de 150 hospitales. Su mediana de edad fue 69,4 años (rango: 18-102 años) y el 57,2% eran hombres. Las prevalencias de hipertensión, dislipemia y diabetes mellitus fueron 50,9%, 39,7% y 19,4%, respectivamente.Los síntomas más frecuentes fueron fiebre (84,2%) y tos (73,5%). Fueron frecuentes los valores elevados de ferritina (73,5%), lactato deshidrogenasa (73,9%) y dímero D (63,8%), así como lalinfopenia (52,8%). Los fármacos antivirales más utilizados fueron la hidroxicloroquina (85,6%)y el lopinavir/ritonavir (61,4%). El 33,1% desarrolló distrés respiratorio. La tasa de mortalidad global fue del 21,0%, con un marcado incremento con la edad (50-59 años: 4,7%; 60-69 años:10,5%; 70-79 años: 26,9%; ≥ 80 años: 46%).Conclusiones: El Registro SEMI-COVID-19 proporciona información sobre las características clínicas de los pacientes con COVID-19 hospitalizados en España. Los pacientes con COVID-19 hospitalizados en España son en su mayoría casos graves, ya que uno de cada 3 pacientes desarrolló distrés respiratorio y uno de cada 5 pacientes falleció. Nuestros datos confirman una estrecha relación entre la edad avanzada y la mortalidadLa Sociedad Española de Medicina Interna (SEMI) es la patrocinadora de este estudio

The Use of Corticosteroids or Tocilizumab in COVID-19 Based on Inflammatory Markers

SEMI-COVID-19 Network.[Background] The inflammatory cascade is the main cause of death in COVID-19 patients. Corticosteroids (CS) and tocilizumab (TCZ) are available to treat this escalation but which patients to administer it remains undefined.[Objective] We aimed to evaluate the efficacy of immunosuppressive/anti-inflammatory therapy in COVID-19, based on the degree of inflammation. .[Design] A retrospective cohort study with data on patients collected and followed up from March 1st, 2020, to May 1st, 2021, from the nationwide Spanish SEMI-COVID-19 Registry. Patients under treatment with CS vs. those under CS plus TCZ were compared. Effectiveness was explored in 3 risk categories (low, intermediate, high) based on lymphocyte count, C-reactive protein (CRP), lactate dehydrogenase (LDH), ferritin, and d-dimer values.[Patients] A total of 21,962 patients were included in the Registry by May 2021. Of these, 5940 met the inclusion criteria for the present study (5332 were treated with CS and 608 with CS plus TCZ).[Main Measures] The primary outcome of the study was in-hospital mortality. Secondary outcomes were the composite variable of in-hospital mortality, requirement for high-flow nasal cannula (HFNC), non-invasive mechanical ventilation (NIMV), invasive mechanical ventilation (IMV), or intensive care unit (ICU) admission.[Key Results] A total of 5940 met the inclusion criteria for the present study (5332 were treated with CS and 608 with CS plus TCZ). No significant differences were observed in either the low/intermediate-risk category (1.5% vs. 7.4%, p=0.175) or the high-risk category (23.1% vs. 20%, p=0.223) after propensity score matching. A statistically significant lower mortality was observed in the very high–risk category (31.9% vs. 23.9%, p=0.049).[Conclusions] The prescription of CS alone or in combination with TCZ should be based on the degrees of inflammation and reserve the CS plus TCZ combination for patients at high and especially very high risk.Peer reviewe