YODA Kinase Controls a Novel Immune Pathway of Tomato Conferring Enhanced Disease Resistance to the Bacterium Pseudomonas syringae

© 2020 Téllez, Muñoz-Barrios, Sopeña-Torres, Martín-Forero, Ortega, Pérez, Sanz, Borja, de Marcos, Nicolas, Jahrmann, Mena, Jordá and Molina.Mitogen-activated protein kinases (MAPK) play pivotal roles in transducing developmental cues and environmental signals into cellular responses through pathways initiated by MAPK kinase kinases (MAP3K). AtYODA is a MAP3K of Arabidopsis thaliana that controls stomatal development and non-canonical immune responses. Arabidopsis plants overexpressing a constitutively active YODA protein (AtCA-YDA) show broad-spectrum disease resistance and constitutive expression of defensive genes. We tested YDA function in crops immunity by heterologously overexpressing AtCA-YDA in Solanum lycopersicum. We found that these tomato AtCA-YDA plants do not show developmental phenotypes and fitness alterations, except a reduction in stomatal index, as reported in Arabidopsis AtCA-YDA plants. Notably, AtCA-YDA tomato plants show enhanced resistance to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 and constitutive upregulation of defense-associated genes, corroborating the functionality of YDA in tomato immunity. This function was further supported by generating CRISPR/Cas9-edited tomato mutants impaired in the closest orthologs of AtYDA [Solyc08g081210 (SlYDA1) and Solyc03g025360 (SlYDA2)]. Slyda1 and Slyda2 mutants are highly susceptible to P. syringae pv. tomato DC3000 in comparison to wild-type plants but only Slyda2 shows altered stomatal index. These results indicate that tomato orthologs have specialized functions and support that YDA also regulates immune responses in tomato and may be a trait for breeding disease resistance.This work was supported by grants BIO2015-64077-R of the Spanish Ministry of Economy and Competitiveness (MINECO), RTI2018-096975-B-I00 of Spanish Ministry of Science, Innovation and Universities, and grant P190050072 of Plant Response Biotech SL to AMo. Research in the Montaña Mena’s laboratory is supported by the MINECO (PPII10-0194-4164) and the Junta de Comunidades de Castilla-La Mancha (SBPLY/17/180501/000394), complemented with EU FEDER funds. JT was financially supported by a PhD fellow of the FPI program from MINECO (BES-2016-076708). AM-F and AO were recipients of research and PhD fellowships from JCCM

ADAM10 gene variants in AD patients and their relationship to CSF protein levels

This article belongs to the Special Issue Advances in the Research of Alzheimer's Disease: From Molecular Basis to Therapy.ADAM10 is the main α-secretase acting in the non-amyloidogenic processing of APP. We hypothesized that certain rare ADAM10 variants could increase the risk for AD by conferring the age-related downregulation of α-secretase. The ADAM10 gene was sequenced in 103 AD cases (82% familial) and 96 cognitively preserved nonagenarians. We examined rare variants (MAF < 0.01) and determined their potential association in the AD group with lower CSF protein levels, as analyzed by means of ELISA, and Western blot (species of 50 kDa, 55 kDa, and 80 kDa). Rare variants were found in 15.5% of AD cases (23% early-onset, 8% late-onset) and in 12.5% of nonagenarians, and some were group-specific. All were intronic variants except Q170H, found in three AD cases and one nonagenarian. The 3′UTR rs74016945 (MAF = 0.01) was found in 6% of the nonagenarians (OR 0.146, p = 0.057). Altogether, ADAM10 total levels or specific species were not significantly different when comparing AD with controls or carriers of rare variants versus non-carriers (except a Q170H carrier exhibiting low levels of all species), and did not differ according to the age at onset or APOE genotype. We conclude that ADAM10 exonic variants are uncommon in AD cases, and the presence of rare intronic variants (more frequent in early-onset cases) is not associated with decreased protein levels in CSF.This work was supported by grants from Instituto de Salud Carlos III (PI20/00469); Ayuda a Neurociencias de la Fundación Tatiana Pérez de Guzmán el Bueno (4564/006); FEDER funds, Spain; and a grant from Direcció General d’Universitat, Investigació i Ciència, GVA (AICO/2018/090). A.G-G is supported by a GVA-Predoctoral fellowship (grant CIACIF/2021/426) from Generalitat Valenciana, Spain.Peer reviewe

MAPK inhibitors dynamically affect melanoma release of immune NKG2D-ligands, as soluble protein and extracellular vesicle-associated

Metastatic melanoma presents, in many cases, oncogenic mutations in BRAF, a MAPK involved in proliferation of tumour cells. BRAF inhibitors, used as therapy in patients with these mutations, often lead to tumour resistance and, thus, the use of MEK inhibitors was introduced in clinics. BRAFi/MEKi, a combination that has modestly increased overall survival in patients, has been proven to differentially affect immune ligands, such as NKG2D-ligands, in drug-sensitive vs. drug-resistant cells. However, the fact that NKG2D-ligands can be released as soluble molecules or in extracellular vesicles represents an additional level of complexity that has not been explored. Here we demonstrate that inhibition of MAPK using MEKi, and the combination of BRAFi with MEKi in vitro, modulates NKG2D-ligands in BRAF-mutant and WT melanoma cells, together with other NK activating ligands. These observations reinforce a role of the immune system in the generation of resistance to directed therapies and support the potential benefit of MAPK inhibition in combination with immunotherapies. Both soluble and EV-associated NKG2D-ligands, generally decreased in BRAF-mutant melanoma cell supernatants after MAPKi in vitro, replicating cell surface expression. Because potential NKG2D-ligand fluctuation during MAPKi treatment could have different consequences for the immune response, a pilot study to measure NKG2D-ligand variation in plasma or serum from metastatic melanoma patients, at different time points during MAPKi treatment, was performed. Not all NKG2D-ligands were equally detected. Further, EV detection did not parallel soluble protein. Altogether, our data confirm the heterogeneity between melanoma lesions, and suggest testing several NKG2D-ligands and other melanoma antigens in serum, both as soluble or vesicle-released proteins, to help classifying immune competence of patients

SARS-CoV-2 Infection in Multiple Sclerosis

To understand COVID-19 characteristics in people with multiple sclerosis (MS) and identify high-risk individuals due to their immunocompromised state resulting from the use of disease-modifying treatments. Retrospective and multicenter registry in patients with MS with suspected or confirmed COVID-19 diagnosis and available disease course (mild = ambulatory; severe = hospitalization; and critical = intensive care unit/death). Cases were analyzed for associations between MS characteristics and COVID-19 course and for identifying risk factors for a fatal outcome. Of the 326 patients analyzed, 120 were cases confirmed by real-time PCR, 34 by a serologic test, and 205 were suspected. Sixty-nine patients (21.3%) developed severe infection, 10 (3%) critical, and 7 (2.1%) died. Ambulatory patients were higher in relapsing MS forms, treated with injectables and oral first-line agents, whereas more severe cases were observed in patients on pulsed immunosuppressors and critical cases among patients with no therapy. Severe and critical infections were more likely to affect older males with comorbidities, with progressive MS forms, a longer disease course, and higher disability. Fifteen of 33 patients treated with rituximab were hospitalized. Four deceased patients have progressive MS, 5 were not receiving MS therapy, and 2 were treated (natalizumab and rituximab). Multivariate analysis showed age (OR 1.09, 95% CI, 1.04-1.17) as the only independent risk factor for a fatal outcome. This study has not demonstrated the presumed critical role of MS therapy in the course of COVID-19 but evidenced that people with MS with advanced age and disease, in progressive course, and those who are more disabled have a higher probability of severe and even fatal diseas

The Caldera. No. 26

“La PALABRA tiene PODER”, más allá del lema que nos convocó en la décima versión del Concurso Inter colegiado de Oratoria, que en el marco de ULIBRO 2023, se llevó a cabo en NEOMUNDO, el 1 de septiembre pasado, esta frase es una profunda convicción que ha movido al Área de Lengua Castellana del Instituto Caldas, a propiciar y mantener este espacio en el que nuestros estudiantes son los protagonistas. Este año, al tratarse de una celebración especial, quisimos que el tema fundamental fuera el inmenso PODER que tiene la PALABRA; la palabra que construye, que transforma, que edifica, que fortalece, que acompaña, que “abriga”, a diario, al ser humano; en un mundo en el que se debe volver a entender y vivenciar, en el hogar, en la escuela, en la sociedad, el poder transformador de vocablos tan importantes como lo son: “Familia, Educación, Vida, Diversidad, Respeto, Literatura, Escritura, Creatividad, Lectura, Inclusión, Paz, Igualdad Social, Democracia, Ciudadanía y Diálogo”, la palabra cobró fuerza en las voces de nuestros educandos; de esta manera, analizar estos temas tiene que ver necesariamente con los “Futuros posibles”, que planteó la Gran Feria del Libro, ULIBRO, en su vigésima primera versión, para que sigamos trabajando por una sociedad mejor, razón de ser de todas las instituciones educativas del mundo entero.“The WORD has POWER”, beyond the motto that summoned us in the tenth version of the Intercollegiate Oratory Contest, which within the framework of ULIBRO 2023, was held in NEOMUNDO, last September 1, this phrase is a deep conviction that has moved the Spanish Language Area of ​​the Caldas Institute to promote and maintain this space in which our students are the protagonists. This year, as it is a special celebration, we wanted the fundamental theme to be the immense POWER that the WORD has; the word that builds, that transforms, that edifies, that strengthens, that accompanies, that “shelters”, daily, the human being; in a world in which we must understand and experience again, at home, at school, in society, the transformative power of words as important as: “Family, Education, Life, Diversity, Respect, Literature, Writing, Creativity, Reading, Inclusion, Peace, Social Equality, Democracy, Citizenship and Dialogue”, the word gained strength in the voices of our students; In this way, analyzing these issues necessarily has to do with the “Possible Futures”, which the Great Book Fair, ULIBRO, proposed in its twenty-first version, so that we continue working for a better society, the reason for being of all institutions. . educational institutions around the world.Modalidad Presencia

Diverse Large HIV-1 Non-subtype B Clusters Are Spreading Among Men Who Have Sex With Men in Spain

In Western Europe, the HIV-1 epidemic among men who have sex with men (MSM) is dominated by subtype B. However, recently, other genetic forms have been reported to circulate in this population, as evidenced by their grouping in clusters predominantly comprising European individuals. Here we describe four large HIV-1 non-subtype B clusters spreading among MSM in Spain. Samples were collected in 9 regions. A pol fragment was amplified from plasma RNA or blood-extracted DNA. Phylogenetic analyses were performed via maximum likelihood, including database sequences of the same genetic forms as the identified clusters. Times and locations of the most recent common ancestors (MRCA) of clusters were estimated with a Bayesian method. Five large non-subtype B clusters associated with MSM were identified. The largest one, of F1 subtype, was reported previously. The other four were of CRF02_AG (CRF02_1; n = 115) and subtypes A1 (A1_1; n = 66), F1 (F1_3; n = 36), and C (C_7; n = 17). Most individuals belonging to them had been diagnosed of HIV-1 infection in the last 10 years. Each cluster comprised viruses from 3 to 8 Spanish regions and also comprised or was related to viruses from other countries: CRF02_1 comprised a Japanese subcluster and viruses from 8 other countries from Western Europe, Asia, and South America; A1_1 comprised viruses from Portugal, United Kingom, and United States, and was related to the A1 strain circulating in Greece, Albania and Cyprus; F1_3 was related to viruses from Romania; and C_7 comprised viruses from Portugal and was related to a virus from Mozambique. A subcluster within CRF02_1 was associated with heterosexual transmission. Near full-length genomes of each cluster were of uniform genetic form. Times of MRCAs of CRF02_1, A1_1, F1_3, and C_7 were estimated around 1986, 1989, 2013, and 1983, respectively. MRCA locations for CRF02_1 and A1_1 were uncertain (however initial expansions in Spain in Madrid and Vigo, respectively, were estimated) and were most probable in Bilbao, Spain, for F1_3 and Portugal for C_7. These results show that the HIV-1 epidemic among MSM in Spain is becoming increasingly diverse through the expansion of diverse non-subtype B clusters, comprising or related to viruses circulating in other countries

Novel PVA–Hyaluronan–Siloxane Hybrid Nanofiber Mats for Bone Tissue Engineering

Hyaluronan (HA) is a natural biodegradable biopolymer; its biological functions include cell adhesion, cell proliferation, and differentiation as well as decreasing inflammation, angiogenesis, and regeneration of damaged tissue. This makes it a suitable candidate for fabricating nanomaterials with potential use in tissue engineering. However, HA nanofiber production is restricted due to the high viscosity, low evaporation rate, and high surface tension of HA solutions. Here, hybrids in the form of continuous and randomly aligned polyvinyl alcohol (PVA)–(HA)–siloxane nanofibers were obtained using an electrospinning process. PVA–HA fibers were crosslinked by a 3D siloxane organic–inorganic matrix via sol-gel that restricts natural hydrophilicity and stiffens the structure. The hybrid nanofiber mats were characterized by FT-IR, micro-Raman spectroscopy, SEM, and biological properties. The PVA/HA ratio influenced the morphology of the hybrid nanofibers. Nanofibers with high PVA content (10PVA-8 and 10PVA-10) form mats with few beaded nanofibers, while those with high HA content (5PVA-8 and 5PVA-10) exhibit mats with mound patterns formed by “ribbon-like” nanofibers. The hybrid nanofibers were used as mats to support osteoblast growth, and they showed outstanding biological properties supporting cell adhesion, cell proliferation, and cell differentiation. Importantly, the 5PVA-8 mats show 3D spherical osteoblast morphology; this suggests the formation of tissue growth. These novel HA-based nanomaterials represent a relevant advance in designing nanofibers with unique properties for potential tissue regeneration

Estrategias para la consolidación en el mercado de Kraftcolor de Macht

Kraftcolor es un rotulador de la empresa alemana Macht, durante los años 2010 y 2011 se detectó que el rotulador Kraftcolor era el producto que menos ventas retribuían a la empresa macht