A Method for Estimation of Population-Densities of Ice Nucleating Active Pseudomonas-Syringae in Buds and Leaves of Mango

Active ice nucleation strains of Pseudomonas syringae pv. syringae have been associated with a necrotic disease in mango trees growing in Málaga (southern Spain). In this paper a simple multiple-tube test is described to estimate the number of active ice nucleation bacteria associated with plant tissues and, also from suspensions of isolated bacterial strains.Peer reviewe

Effects of ispaghula husk and guar gum on postprandial glucose and insulin concentrations in healthy subjects

P. 235-243The aim of this study was to evaluate, under the same experimental conditions and in the same subjects, the effects of ispaghula husk and guar gum on postprandial glucose and insulin concentrations in healthy female subjects(ten healthy female volunteers aged 30 ± 48 y with normal body mass indices).S

The GB4.0 Platform, an All-In-One Tool for CRISPR/Cas-Based Multiplex Genome Engineering in Plants

CRISPR/Cas ability to target several loci simultaneously (multiplexing) is a game-changer in plant breeding. Multiplexing not only accelerates trait pyramiding but also can unveil traits hidden by functional redundancy. Furthermore, multiplexing enhances dCas-based programmable gene expression and enables cascade-like gene regulation. However, the design and assembly of multiplex constructs comprising tandemly arrayed guide RNAs (gRNAs) requires scarless cloning and is still troublesome due to the presence of repetitive sequences, thus hampering a more widespread use. Here we present a comprehensive extension of the software-assisted cloning platform GoldenBraid (GB), in which, on top of its multigene cloning software, we integrate new tools for the Type IIS-based easy and rapid assembly of up to six tandemly-arrayed gRNAs with both Cas9 and Cas12a, using the gRNA-tRNA-spaced and the crRNA unspaced approaches, respectively. As stress tests for the new tools, we assembled and used for Agrobacterium-mediated stable transformation a 17 Cas9-gRNAs construct targeting a subset of the Squamosa-Promoter Binding Protein-Like (SPL) gene family in Nicotiana tabacum. The 14 selected genes are targets of miR156, thus potentially playing an important role in juvenile-to-adult and vegetative-to-reproductive phase transitions. With the 17 gRNAs construct we generated a collection of Cas9-free SPL edited T plants harboring up to 9 biallelic mutations and showing leaf juvenility and more branching. The functionality of GB-assembled dCas9 and dCas12a-based CRISPR/Cas activators and repressors using single and multiplexing gRNAs was validated using a Luciferase reporter with the Solanum lycopersicum Mtb promoter or the Agrobacterium tumefaciens nopaline synthase promoter in transient expression in Nicotiana benthamiana. With the incorporation of the new web-based tools and the accompanying collection of DNA parts, the GB4.0 genome edition turns an all-in-one open platform for plant genome engineering

Cat epididymal semen cryopreserved with and without vitamin E: effect on sperm parameters and lipid peroxidation

The aims of this study were to investigate: 1) if the addition of \u3b1-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of \u3b1-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) \u2013 epididymal sperm were frozen with a commercial Botucrio\uae extender; group 0.3, group 0.6 and group 0.9 \u2013 the extender was supplemented with 0.3, 0.6 and 0.9 mM of \u3b1-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three \u3b1-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the \u3b1-tocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of \u3b1-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of \u3b1-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages

Severe manifestations of SARS-CoV-2 in children and adolescents: from COVID-19 pneumonia to multisystem inflammatory syndrome: a multicentre study in pediatric intensive care units in Spain

Background Multisystem inflammatory syndrome temporally associated with COVID-19 (MIS-C) has been described as a novel and often severe presentation of SARS-CoV-2 infection in children. We aimed to describe the characteristics of children admitted to Pediatric Intensive Care Units (PICUs) presenting with MIS-C in comparison with those admitted with SARS-CoV-2 infection with other features such as COVID-19 pneumonia. Methods A multicentric prospective national registry including 47 PICUs was carried out. Data from children admitted with confirmed SARS-CoV-2 infection or fulfilling MIS-C criteria (with or without SARS-CoV-2 PCR confirmation) were collected. Clinical, laboratory and therapeutic features between MIS-C and non-MIS-C patients were compared. Results Seventy-four children were recruited. Sixty-one percent met MIS-C definition. MIS-C patients were older than non-MIS-C patients (p = 0.002): 9.4 years (IQR 5.5–11.8) vs 3.4 years (IQR 0.4–9.4). A higher proportion of them had no previous medical history of interest (88.2% vs 51.7%, p = 0.005). Non-MIS-C patients presented more frequently with respiratory distress (60.7% vs 13.3%, p < 0.001). MIS-C patients showed higher prevalence of fever (95.6% vs 64.3%, p < 0.001), diarrhea (66.7% vs 11.5%, p < 0.001), vomits (71.1% vs 23.1%, p = 0.001), fatigue (65.9% vs 36%, p = 0.016), shock (84.4% vs 13.8%, p < 0.001) and cardiac dysfunction (53.3% vs 10.3%, p = 0.001). MIS-C group had a lower lymphocyte count (p < 0.001) and LDH (p = 0.001) but higher neutrophil count (p = 0.045), neutrophil/lymphocyte ratio (p < 0.001), C-reactive protein (p < 0.001) and procalcitonin (p < 0.001). Patients in the MIS-C group were less likely to receive invasive ventilation (13.3% vs 41.4%, p = 0.005) but were more often treated with vasoactive drugs (66.7% vs 24.1%, p < 0.001), corticosteroids (80% vs 44.8%, p = 0.003) and immunoglobulins (51.1% vs 6.9%, p < 0.001). Most patients were discharged from PICU by the end of data collection with a median length of stay of 5 days (IQR 2.5–8 days) in the MIS-C group. Three patients died, none of them belonged to the MIS-C group. Conclusions MIS-C seems to be the most frequent presentation among critically ill children with SARS-CoV-2 infection. MIS-C patients are older and usually healthy. They show a higher prevalence of gastrointestinal symptoms and shock and are more likely to receive vasoactive drugs and immunomodulators and less likely to need mechanical ventilation than non-MIS-C patients

Revista de Vertebrados de la Estación Biológica de Doñana

La reproducción de Hyla meridionalis en el suroeste de EspañaAlimentación y relaciones tróficas entre larvas de Triturus marmoratus, T. alpestris y T. helveticus (Amphibia: CaudataOrganization of behaviour in isolated lizards (Gallotia galloti galloti) as revealed by multivariate analyseComposición y estructura de las comunidades de aves a lo largo de un gradiente altitudinal en tres medios arbustivos del macizo de Ayllón(Sistema Central)La comunidad de aves de un acebuchar del sur de España durante el periodo invernal y de cria.Alimentación del buho chico (Asio otus) en la isla de Tenerife y análisis comparativo con la dieta de Tyto albaAlimentación del zorzal charlo (Turdus viscivoros) en la sierra de Cazorla, SE de España.La migración en España del verderón común (Carduelis chloris, L.) según los resultados de anillamientoIdentificación de los principales quirópteros ibéricos a partir de sus dientes aislados. Valor sistemático de los caracteres morfológicos y métricos dentariosRitmo de actividad en Gazella dorcasSobre la sistemática y biología de Eliomys quercinus en la Cordillera Cantábrica.Primeras citas de Barbus microcephalus Almaça, 1967 (Ostariophysi: Cyprinidae) en EspañaSobre la presencia de Lampetra planeri BLOCH, 1784 en España.Sobre el status taxonómico del género Valencia Myers, 1928 en el suroeste de IberiaNuevas localidades de Chondrostroma polypis Steindachner, 1865 (Ostariophysi, Cyprinidae) en España.Sobre la distribución Gobio gobio (L., 1758)(OSTAOPHYSI, CIPRINIDAE) en EspañaNotas sobre la alimentación de larvas de anfibios: 2. Salamandra salamandra de CazorlaNuevos datos sobre la permanencia de caracteres larvarios en individuos adultos de una población de tritón pirenaico (Euproctus asper) en el valle de AránLa variación del diseño natural como método de reconocimiento individual en Triturus boscaiPresencia de Triturus boscai en la provincia de Córdoba.Datos sobre la alimentación de Athene cunicularia en la Reserva de fauna altoandina de Ulla-Ulla, Bolivia.Falco peregrinus cassini en BoliviaAlgunos datos sobre quirópteros de Galicia.Notas sobre la alimentación de la nutria (Lutra lutra) en el embalse de Matavacas, HuelvaPeer reviewe