The role of two families of bacterial enzymes in putrescine synthesis from agmatine via agmatine deiminase

Putrescine, one of the main biogenic amines associated to microbial food spoilage, can be formed by bacteriafrom arginine via ornithine decarboxylase (ODC), or from agmatine via agmatine deiminase (AgDI). This study aims to correlate putrescine production from agmatine to the pathway involving N-carbamoylputrescine formation via AdDI (the aguAproduct) and N-carbamoylputrescine amidohydrolase (the aguB product), or putrescine carbamoyltransferase (the ptcA product) in bacteria. PCR methods were developed to detect the two genes involved in putrescine production from agmatine.Putrescine production from agmatine could be linked to the  aguA and  ptcA genes in  Lactobacillus hilgardii X1B,Enterococcus faecalis ATCC 11700, and Bacillus cereus ATCC 14579. By contrast Lactobacillus sakei 23K was unable toproduce putrescine, and although a fragment of DNA corresponding to the gene aguA was amplified, no amplification wasobserved for the ptcA gene. Pseudomonasaeruginosa PAO1 produces putrescine and is reported to harbour aguA and aguBgenes, responsible for agmatine deiminase and N-carbamoylputrescine amidohydrolase activities. The enzyme from P. aeruginosa PAO1 that converts N-carbamoylputrescine to putrescine (the aguB product) is different from other microorganismsstudied (the ptcA product). Therefore, the aguB gene from P. aeruginosa PAO1 could not be amplified with ptcA specificprimers. The aguB and ptcA genes have frequently been erroneously annotated in the past, as in fact these two enzymes areneither homologous nor analogous. Furthermore, the aguA, aguB and ptcA sequences available from GenBank were subjected to phylogenetic analysis, revealing that gram-positive bacteria harboured ptcA, whereas gram-negative bacteria harbouraguB. This paper also discusses the role of the agmatine deiminase system (AgDS) in acid stress resistance.&nbsp

Insecticidal activities of diketopiperazines of Nomuraea rileyi entomopathogenic fungus

Entomopathogenic fungi are fungal organisms extensively used in various parts of the world as biopesticides against insect pests that cause important economic damage. Various secondary metabolites produced by these fungi have many potential biological activities. The present study was undertaken to evaluate the insecticidal activity of extracts and pure compounds from Nomuraea rileyi (Farlow) Samson entomopathogenic fungi against Spodoptera frugiperda Smith (Lepidoptera), Ceratitis capitata Wiedemann (Diptera) and Tribolium castaneum Herbst (Coleoptera), three insect pests that generate serious economic losses in the northwest of Argentina. Diketopiperazines were extracted from the culture free supernatant of the media with ethyl acetate. Antifeedant properties were detected in all extracts under dietary choice conditions (300 ug/ g of diet). The maximum antifeedant activity was noted in cycles (Pro-Val) (86.02) and cycle (Pro-Phe) (73.47), while the rest of the extracts and metabolites exhibited varying degrees of moderate or less toxic effects. The maximum oviposition deterrence against C. capitata (55.86%) was recorded with cycle (Pro-Phe) at a 50 µm/cm2 dose. Culture medium extracts supplemented with insect remains and all pure compounds showed repellent action against T. castaneum. The main repellency was observed in phenylacetic acid and cycle (Pro-Val) with RI values of 42 and 41% respectively. The present study would suggest the possible utilization of entomopathogenic fungal metabolites as an effective agent for controlling insect pests that cause important economic losses

Synthesis, structure, and biological assays of novel trifluoromethyldiazepine–metal complexes

A new series of CuII, NiII, CoII, and MnIII complexes have been synthesised from the (6Z)-6-(7-trifluoromethyl-1,2,3,4-tetrahydro-5H-1,4-diazepin-5-ylidene)cyclohexa-2,4-dien-1-one (HDZP) ligand. These complexes were characterised by elemental, spectroscopic (IR and UV-vis), and thermal analysis. The crystal structure of Cu-DZP was solved by X-ray diffraction methods. The complex crystallises in the monoclinic P21/c space group, with two molecules per unit cell. The crystal lattice is stabilised by different intra and intermolecular interactions. Hirshfeld surface analysis was employed to obtain additional information about interactions that are responsible for the crystal packing. Quantitative examination of the fingerprint plots indicated the dominant contribution of H⋯H and H⋯X (X = O, F) interactions in the crystal packing. In addition, C–H⋯chelate ring (CR) and C–H⋯π interactions are described in detail and evaluated using DFT calculations. The antibacterial properties and the mechanism of inhibition of the main bacterial resistant mechanism, the biofilm, of the metal complexes and free ligand were investigated. [Mn(DZP)₃]·2H₂O was the most active complex against the Pseudomonas aeruginosa biofilm formation with an inhibition of 40 %. However, none of the complexes inhibit more than 25 % of the Gram negative bacteria microbial development. The most meaningful result was the bactericidal effect of [Co(DZP)₂(H₂O)₂]·2H₂O against the Gram positive bacteria, Staphylococcus aureus, which inhibits the bacterial development and significantly reduces the biofilm formation at low concentration.Instituto de Física La PlataPlanta Piloto Multipropósito - Laboratorio de Servicios a la Industria y al Sistema CientíficoCentro de Química Inorgánic

Subproductos de la Industria Vitivinícola de los Valles Calchaquíes como Fuente de Metabolitos Bioactivos

Uno de los principales desafíos de nuestra sociedad es el desarrollo de soluciones sostenibles para la gestión de los subproductos y desechos de agroindustrias. Numerosos estudios han demostrado que los subproductos del procesamiento de alimentos son fuentes ricas en fibras dietéticas, metabolitos secundarios, vitaminas, proteínas y péptidos y que pueden usarse como ingredientes alimenticios naturales o como nutracéuticos de bajo costo (Schieber, 2019). La industria vitivinícola genera una gran cantidad de residuos orgánicos que resultan, altamente, contaminantes para el medio ambiente. Al ser el cultivo de la vid un cultivo estacional, la producción de vino se restringe a pocos meses, produciendo una gran cantidad de desechos con elevada carga de materia orgánica en poco tiempo. Argentina es el quinto productor mundial de vino y dependiendo de las condiciones de cosechas de las uvas, los residuos pueden alcanzar el 20% del volumen total. El orujo que constituye el 62% de los residuos generados en la bodega se produce durante el prensado de la uva y está constituidos principalmente por piel y semillas de la baya. Por lo general, estos residuos son quemados, usados para la alimentación del ganado, como abono o desechados en ríos, a pesar de que contienen fitocompuestos atractivos para las industrias farmacéutica, cosmética y alimentaria (García-Lomillo et al., 2017).Entre los fitoquímicos de uvas vinculados a efectos beneficiosos en salud, se incluyen: alcaloides, terpenos, saponinas, aceites volátiles y un amplio y diverso grupo de compuestos fenólicos (ác. fenólicos, flavonoides antocianos, flavonas, flavanoles, estilbenos, taninos, etc.) abundantes en la piel y semillas de las uvas (Ananga et al., 2017). Los flavonoides tienen efectos antivirales, anticancerígenos, antioxidantes, antimicrobiano, antiinflamatorio, anticolesterolémicos, antiangiogénicos y antitrombogénicos (Teixeira et al., 2014; Lingua et al., 2016; Ananga et al., 2017; Mattos et al., 2017). Otro beneficio es que, los compuestos fenólicos de vino que no son absorbidos en el intestino delgado llegan al colon donde son fermentados y desconjugados por enzimas bacterianas y son capaces de inhibir el crecimiento de bacterias potencialmente patógenas, suprimir factores de virulencias como la neutralización de toxinas bacterianas, inhibir la formación de biofilm, y reducir de adherencia bacteriana (Vázquez-Armenta et al., 2018).En los últimos años, los polifenoles han atraído un creciente interés por sus beneficios potenciales para la salud en la prevención de enfermedades cardíacas, hepáticas, neurodegenerativas, respiratorias, intestinales, síndrome metabólico y ciertos tipos de cáncer, por lo que se utilizan en diferentes productos alimenticios (como colorantes o antioxidantes alimenticios) y en aplicaciones farmacéuticas (como nutracéuticos) (Georgiev et al., 2014; Schieber, 2019). Sin embargo, el perfil polifenólico se ve fuertemente afectado tanto cuantitativa como cualitativamente por la variedad de uva, su grado de madurez, el origen geográfico, el clima, las condiciones del suelo del viñedo y técnicas de vinificación (Jiang & Zhang 2018; Giovinazzo et al., 2020).Los Valles Calchaquíes ubicados en la región noroeste de Argentina tienen un clima templado con notables amplitudes térmicas y ocasionalmente presentan heladas tardías prolongadas en primavera. Los vinos blancos regionales corresponden al varietal Torrontés, que se ha convertido en la variedad emblemática de la región. Esta uva es óptima para producir vinos aromáticos y se adapta muy bien a toda la zona, convirtiéndose en la más cultivada de la región (Instituto Nacional De Vitivinicultura, 2021). Por otro lado, el vino tinto varietal Malbec (variedad de uva morada más explotada de Argentina) de los Valles Calchaquíes presenta particularidades diferentes a otras regiones vitivinícolas del país debido a las características del terruño descritas anteriormente. En vista de lo anterior, se decidió estudiar la composición química y propiedades antioxidante, antipatogénicas, citotoxicidad, capacidad de inhibición enzimática y toxicidad aguda de orujos Torrontés y Malbec (Vitis vinifera L.) de los Valles Calchaquíes (Argentina). Para este estudio el orujo de uva Torrontés se obtuvo de un proceso de elaboración de vino blanco, lo que significa que las uvas no fueron sometidas a fermentación etanólica, a diferencia de lo que ocurre en el proceso de elaboración de vino tinto, donde las uvas están totalmente involucradas en la fermentación como seria en el caso del orujo Malbec. El porcentaje de humedad en los orujos de Malbec y Torrontés fue de 73,6 y 74,4%, respectivamente. Después de secar y moler en harina, los orujos de uva se sometieron a extracción utilizando solventes GRAS (etanol/agua) y se llevaron a sequedad. Los rendimientos de extracción de principios solubles del orujo de Malbec y Torrontés fueron del 14 y 10%, respectivamente. Se caracterización química (tamizaje fitoquímico) de los extractos de orujo ha permitido identificar varios metabolitos secundarios entre ellos polifenoles, flavonoides, antocianinas, cumarinas, taninos, quinonas y terpenos. Los esteroles se evidencian únicamente en el orujo de Torrontés, mientras que ninguno de los extractos presenta alcaloides y saponinas. El análisis cuantitativo de los metabolitos fenólicos presentes en los extractos revela que la variedad tinta Malbec presenta 6,7 veces más contenido de fenoles totales que el Torrontés. El contenido de compuestos fenólicos no flavonoides representa el 20% de los polifenoles presentes en el extracto de orujo de Malbec y el 40% en el orujo Torrontés. Respecto a los compuestos flavonoides totales el extracto Malbec presenta 7,5 veces más que el Torrontés. Esto se debe a que el contenido de flavanonas/dihidroflavonoles en el extracto de Malbec es 6,3 veces superior al del Torrontés y que en este último carece de antocianinas presentes en el orujo de vino tinto. Asimismo, el contenido de favonas/flavonoles en el extracto Torrontés (0,1%) es muy inferior al Malbec (7,4%). El contenido de taninos en ambos extractos es significativo.Mediante el análisis HPLC-DAD de los extractos se identifican 35 compuestos fenólicos individuales pertenecientes a la familia de los ácidos fenólicos, flavonoides y estilbenos. En el extracto Torrontés, los principales polifenoles son: ácido 4,5-di-O-cafeoilquínico ˃ácido 4-O-cafeoilquínico ˃ kaempferol-3-O-glucósido ˃ isorhamnetin-3-O-glucósido ˃ ácido caftárico ˃ ácido sinápico ˃ ácido gálico ˃ ácido protocatéquico ˃ ácido trans-ferúlico ˃ (+)-catequina. Mientras que en el extracto Malbec los principales compuestos fenólicos son: ácido protocatequico ˃ (+)-catequina ˃ ácido gálico ˃ ácido 4-O-cafeoilquínico ˃ ácido sinápico. El contenido de ácido 4,5-di-O-cafeoilquínico en el extracto Torrontés es 5 veces superior al Malbec. Mientras que, el contenido de (+)-catequina y de los ácidos protocatéquico y gálico es 8,8; 10 y 2,7 veces superior en el extracto Malbec.Las especies reactivas, particularmente EROs (especies reactivas centradas en átomo de oxígeno) y ERNs (especies reactivas centradas en átomo de nitrógeno), juegan un papel importante en varios procesos fisiológicos, a saber, señalización celular, cascada inflamatoria y homeostasis. De esta forma, la evaluación de la capacidad depuradora de un extracto contra EROs y ERNs se vuelve más interesantes debido a sus funciones claves en los tejidos vivos. Con respecto las actividades antiradical catión ABTS (CI50=7,79 ± 0,17 µg/ml), óxido nítrico (CI50=414,19 ± 5,79 µg/ml), anión superóxido (CI50=74,17 ± 4,12 µg/ml), e hipoclorito (CI50=6,71 ± 0,36 µg/ml) ensayadas, el extracto Malbec muestra la mayor eficiencia de depuración. En el ensayo del poder quelante de hierro, el extracto Malbec es capaz de quelar el 42% del metal a 1000 µg/ml. Asimismo el extracto Malbec (CR50= 10,22 ± 0,16 µg/ml) presenta poder reductor del Fe3+ superior al extracto Torrontés (CR50= 84,62 ± 0,95 µg/ml). En el ensayo de capacidad antioxidante in vivo que utiliza la levadura Saccharomyces cerevisiae para estudiar la respuesta celular a EROs, el extracto de orujo Malbec (12,5 µg/ml) rescata el 24% de la levadura del estrés oxidativo inducido por H2O2. En síntesis, el orujo Malbec muestra buena actividad depuradora de ROS y RNS, lo que puede estar relacionado con su mayor contenido fenólico respecto al orujo Torrontés, particularmente catequina, ácido gálico y ácido protocatequico cuyo potencial depurativo de radicales libres ha sido ampliamente demostrado.En diferentes bioensayos con dos líneas celulares de adenocarcinoma colorrectal humano se analiza la citotoxicidad de estos extractos. El orujo Torrontés (1 mg/ml) afecta la viabilidad del 70 y 50% de las células HT29-MTX y Caco-2, respectivamente. Mientras que el orujo Malbec a la misma concentración reduce el 20% de la actividad metabólica de las células Caco-2.La tirosinasa, también conocida como polifenol oxidasa, es la enzima clave en la producción de melanina. Inhibidores de esta enzima tienen gran interés en productos médicos y cosméticos ya que pueden usarse para prevenir o tratar los problemas de hiperpigmentación. El extracto de orujo Malbec inhibe la actividad de esta enzima (IC50= 88,8 ± 2,36 µg/ml). Sin embargo, ningún extracto inhibió la enzima xantina oxidasa que desempeña un papel clave en la hiperuricemia.Por otro lado, los extractos de orujo presentan actividad antipatogénica ya que inhiben la producción de la biopelícula bacteriana, principal causa de contaminación en las industrias de alimentos; y la actividad metabólica de Pseudomonas aeruginosa y Staphylococcus aureus en el entorno del biofilm. Existe una correlación positiva entre la actividad antibiofilm y antioxidante y el contenido de polifenoles de los extractos más activos (Viola et al., 2018, 2021). La atenuación de la biopelícula y de la motilidad swarming y swimming responsable del desplazamiento de P. aeruginosa por acción del extracto de Torrontés, está controlada por quorum sensing (Viola et al., 2020).El ensayo de toxicidad aguda con Artemia salina es un método in vivo simple y rápido que permite evaluar la citotoxicidad de una muestra y se correlaciona bien con los ensayos in vivo. Las concentraciones biológicamente activas de los extractos de orujo no son letales para el crustáceo sugiriendo su uso seguro.Los contenidos de metabolitos fenólicos y actividad biológica encontrados en los orujos de los Valles Calchaquíes les confieren propiedades promotoras de la salud humana, con la correspondiente revalorización de los residuos de las bodegas regionales.Fil: Tapia, Pablo Ezequiel. Universidad Nacional de Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria; ArgentinaFil: Silva, Ana M.. Polytechnic of Porto; PortugalFil: Delerue Matos, Cristina. Polytechnic of Porto; PortugalFil: Moreira, Manuela. Polytechnic of Porto; PortugalFil: Rodriguez, Francisca. Polytechnic of Porto; PortugalFil: Ortega, María Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Santi, María Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Viola, Carolina Maria. Universidad Nacional de Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria; ArgentinaFil: Torres Carro, Romina. Universidad Nacional de Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria; ArgentinaFil: Cartagena, Elena. Universidad Nacional de Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria; ArgentinaFil: Arena, Mario Eduardo. Universidad Nacional de Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria; ArgentinaFil: Alberto, Maria Rosa. Universidad Nacional de Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Biotecnología Farmacéutica y Alimentaria; Argentina2do Taller de Biotecnología Aplicada a la Tecnología de AlimentosCiudad Autónoma de Buenos AriesArgentinaUniversidad Tecnológica Nacional. Facultad Regional Buenos AiresMinisterio de Ciencia, Tecnología e Innovación. Agencia Nacional de Promoción Científica y Tecnológic

A chemical survey of exoplanets with ARIEL

Thousands of exoplanets have now been discovered with a huge range of masses, sizes and orbits: from rocky Earth-like planets to large gas giants grazing the surface of their host star. However, the essential nature of these exoplanets remains largely mysterious: there is no known, discernible pattern linking the presence, size, or orbital parameters of a planet to the nature of its parent star. We have little idea whether the chemistry of a planet is linked to its formation environment, or whether the type of host star drives the physics and chemistry of the planet’s birth, and evolution. ARIEL was conceived to observe a large number (~1000) of transiting planets for statistical understanding, including gas giants, Neptunes, super-Earths and Earth-size planets around a range of host star types using transit spectroscopy in the 1.25–7.8 μm spectral range and multiple narrow-band photometry in the optical. ARIEL will focus on warm and hot planets to take advantage of their well-mixed atmospheres which should show minimal condensation and sequestration of high-Z materials compared to their colder Solar System siblings. Said warm and hot atmospheres are expected to be more representative of the planetary bulk composition. Observations of these warm/hot exoplanets, and in particular of their elemental composition (especially C, O, N, S, Si), will allow the understanding of the early stages of planetary and atmospheric formation during the nebular phase and the following few million years. ARIEL will thus provide a representative picture of the chemical nature of the exoplanets and relate this directly to the type and chemical environment of the host star. ARIEL is designed as a dedicated survey mission for combined-light spectroscopy, capable of observing a large and well-defined planet sample within its 4-year mission lifetime. Transit, eclipse and phase-curve spectroscopy methods, whereby the signal from the star and planet are differentiated using knowledge of the planetary ephemerides, allow us to measure atmospheric signals from the planet at levels of 10–100 part per million (ppm) relative to the star and, given the bright nature of targets, also allows more sophisticated techniques, such as eclipse mapping, to give a deeper insight into the nature of the atmosphere. These types of observations require a stable payload and satellite platform with broad, instantaneous wavelength coverage to detect many molecular species, probe the thermal structure, identify clouds and monitor the stellar activity. The wavelength range proposed covers all the expected major atmospheric gases from e.g. H2O, CO2, CH4 NH3, HCN, H2S through to the more exotic metallic compounds, such as TiO, VO, and condensed species. Simulations of ARIEL performance in conducting exoplanet surveys have been performed – using conservative estimates of mission performance and a full model of all significant noise sources in the measurement – using a list of potential ARIEL targets that incorporates the latest available exoplanet statistics. The conclusion at the end of the Phase A study, is that ARIEL – in line with the stated mission objectives – will be able to observe about 1000 exoplanets depending on the details of the adopted survey strategy, thus confirming the feasibility of the main science objectives.Peer reviewedFinal Published versio

Etest® versus broth microdilution for ceftaroline MIC determination with Staphylococcus aureus: results from PREMIUM, a European multicentre study

Objectives: To compare the concordance of ceftaroline MIC values 24 by reference broth microdilution (BMD) and Etest (BioMérieux, France) for MSSA and MRSA isolates, respectively, in isolates from PREMIUM (D372SL00001), a European multi-centre study.  Methods: Ceftaroline MICs were determined by reference BMD and by Etest for 1,242 MSSA and MRSA from adult patients with community-acquired pneumonia or complicated skin and soft tissue infections collected between February and May 2012; tests were performed across six European laboratories. Selected isolates with ceftaroline resistance in broth (MIC >1 mg/L) were retested in three central laboratories to confirm their behaviour.  Results: Overall concordance between BMD and Etest was good, with >97% essential agreement and >95% categorical agreement. Nevertheless, 12 of the 26 MRSA isolates found resistant by BMD scored as susceptible by Etest, with MICs ≤1 mg/L, thus counting as very major errors, whereas only five of 380 MRSA found ceftaroline susceptible in BMD were mis-categorised as resistant by Etest. Twenty-one of the 26 isolates with MICs of 2 mg/L by BMD were then re-tested twice by each of three central laboratories: BMD MICs of 2 mg/L were consistently found for 19 of the 21 isolates. Among 147 Etest results for these 21 isolates (original plus six repeats per isolate) 112 were >1 mg/L.  Conclusions: BMD and Etest have good overall agreement for ceftaroline against Staphylococcus aureus; nevertheless, reliable Etest-based discrimination of the minority of ceftaroline-resistant (MIC 2 mg/L) MRSA is extremely challenging, requiring careful reading of strips, ideally with duplicate testing

Disease-Modifying Therapies and Coronavirus Disease 2019 Severity in Multiple Sclerosis

Objective: This study was undertaken to assess the impact of immunosuppressive and immunomodulatory therapies on the severity of coronavirus disease 2019 (COVID-19) in people with multiple sclerosis (PwMS). Methods: We retrospectively collected data of PwMS with suspected or confirmed COVID-19. All the patients had complete follow-up to death or recovery. Severe COVID-19 was defined by a 3-level variable: mild disease not requiring hospitalization versus pneumonia or hospitalization versus intensive care unit (ICU) admission or death. We evaluated baseline characteristics and MS therapies associated with severe COVID-19 by multivariate and propensity score (PS)-weighted ordinal logistic models. Sensitivity analyses were run to confirm the results. Results: Of 844 PwMS with suspected (n = 565) or confirmed (n = 279) COVID-19, 13 (1.54%) died; 11 of them were in a progressive MS phase, and 8 were without any therapy. Thirty-eight (4.5%) were admitted to an ICU; 99 (11.7%) had radiologically documented pneumonia; 96 (11.4%) were hospitalized. After adjusting for region, age, sex, progressive MS course, Expanded Disability Status Scale, disease duration, body mass index, comorbidities, and recent methylprednisolone use, therapy with an anti-CD20 agent (ocrelizumab or rituximab) was significantly associated (odds ratio [OR] = 2.37, 95% confidence interval [CI] = 1.18-4.74, p = 0.015) with increased risk of severe COVID-19. Recent use (<1 month) of methylprednisolone was also associated with a worse outcome (OR = 5.24, 95% CI = 2.20-12.53, p = 0.001). Results were confirmed by the PS-weighted analysis and by all the sensitivity analyses. Interpretation: This study showed an acceptable level of safety of therapies with a broad array of mechanisms of action. However, some specific elements of risk emerged. These will need to be considered while the COVID-19 pandemic persists

COVID-19 Severity in Multiple Sclerosis: Putting Data Into Context

Background and objectives: It is unclear how multiple sclerosis (MS) affects the severity of COVID-19. The aim of this study is to compare COVID-19-related outcomes collected in an Italian cohort of patients with MS with the outcomes expected in the age- and sex-matched Italian population. Methods: Hospitalization, intensive care unit (ICU) admission, and death after COVID-19 diagnosis of 1,362 patients with MS were compared with the age- and sex-matched Italian population in a retrospective observational case-cohort study with population-based control. The observed vs the expected events were compared in the whole MS cohort and in different subgroups (higher risk: Expanded Disability Status Scale [EDSS] score > 3 or at least 1 comorbidity, lower risk: EDSS score ≤ 3 and no comorbidities) by the χ2 test, and the risk excess was quantified by risk ratios (RRs). Results: The risk of severe events was about twice the risk in the age- and sex-matched Italian population: RR = 2.12 for hospitalization (p < 0.001), RR = 2.19 for ICU admission (p < 0.001), and RR = 2.43 for death (p < 0.001). The excess of risk was confined to the higher-risk group (n = 553). In lower-risk patients (n = 809), the rate of events was close to that of the Italian age- and sex-matched population (RR = 1.12 for hospitalization, RR = 1.52 for ICU admission, and RR = 1.19 for death). In the lower-risk group, an increased hospitalization risk was detected in patients on anti-CD20 (RR = 3.03, p = 0.005), whereas a decrease was detected in patients on interferon (0 observed vs 4 expected events, p = 0.04). Discussion: Overall, the MS cohort had a risk of severe events that is twice the risk than the age- and sex-matched Italian population. This excess of risk is mainly explained by the EDSS score and comorbidities, whereas a residual increase of hospitalization risk was observed in patients on anti-CD20 therapies and a decrease in people on interferon

DMTs and Covid-19 severity in MS: a pooled analysis from Italy and France

We evaluated the effect of DMTs on Covid-19 severity in patients with MS, with a pooled-analysis of two large cohorts from Italy and France. The association of baseline characteristics and DMTs with Covid-19 severity was assessed by multivariate ordinal-logistic models and pooled by a fixed-effect meta-analysis. 1066 patients with MS from Italy and 721 from France were included. In the multivariate model, anti-CD20 therapies were significantly associated (OR = 2.05, 95%CI = 1.39–3.02, p < 0.001) with Covid-19 severity, whereas interferon indicated a decreased risk (OR = 0.42, 95%CI = 0.18–0.99, p = 0.047). This pooled-analysis confirms an increased risk of severe Covid-19 in patients on anti-CD20 therapies and supports the protective role of interferon