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Prefoldins Negatively Regulate Cold Acclimation in Arabidopsis thaliana by Promoting Nuclear Proteasome-Mediated HY5 Degradation

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Molecular Plant 2017年第6期10卷 791-804页

作者： Carlos Perea-Resa miguel A. Rodriguez-Milla Elisa Iniesto Vicente rubio Julio Salinas Departamento de Biologia Medioambiental Centro de Investigaciones Biologicas CSIC 28040 Madrid Spain Departamento de Genetica Molecular de Plantas Centro Nacional de Biotecnologia CSIC 28049 Madrid Spain present address: Department of Molecular Biology Massachusetts General Hospital Boston MA 02214 USA Present address: Unidad de Biotecnologia Celular Instituto de Salud Garlos Ⅲ 28222 Madrid Spain

The process of cold acclimation is an important adaptive response whereby many plants from temperate regions increase their freezing tolerance after being exposed to low non-freezing temperatures. The correct development of this response relies on proper accumulation of a number of transcription factors that regulate expression patterns of cold-responsive genes. Multiple studies have revealed a variety of molecular mechanisms involved in promoting the accumulation of these transcription factors. Interestingly, however, the mechanisms implicated in controlling such accumulation to ensure their adequate levels remain largely unknown. In this work, we demonstrate that prefoldins （PFDs） control the levels of HY5, an Arabidopsis transcription factor with a key role in cold acclimation by activating anthocyanin biosynthesis, in response to low temperature. Our results show that, under cold conditions, PFDs accumulate into the nucleus through a DELLA-dependent mechanism, where they interact with HY5, triggering its ubiquitination and subsequent degradation. The degradation of HY5 would result, in turn, in anthocyanin biosynthesis attenuation, ensuring the accurate development of cold acclimation. These findings uncover an unanticipated nuclear function for PFDs in plant responses to abiotic stresses.

关键词： prefoldins cold acclimation Arabidopsis HY5 anthocyanin DELLAs

Nested multiplex PCR for identification and detection of human Plasmodium species including Plasmodium knowlesi

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Asian Pacific Journal of Tropical Medicine 2017年第3期10卷 280-284页

作者： Maria miguel-Oteo Adela I Jiram Thuy H Ta-Tang Marta Lanza Shamilah Hisam José miguel rubio Malaria & Emerging Parasitic Diseases Laboratory Parasitology DepartmentNational Microbiology Centre. Instituto de Salud Carlos Ⅲ Majadahonda Unit Institut Penyelidikan PerubatanInstitute for Medical Research

Objective:To develop a new technique for diagnosis of Plasmodium knowlesi and at the same time to be able to discriminate among the diverse species of Plasmodium causing human malaria.Methods:In this study the nested multiplex malaria PCR was redesigned,targeting the 18S rR NA gene,to identify the fifth human Plasmodium species,Plasmodium knowlesi,together with the other human Plasmodium(Plasmodium falciparum,Plasmodium vivax,Plasmodium ovale and Plasmodium malariae)by amplified fragment size using only two amplification processes and including an internal reaction control to avoid false negatives.Results:The technique was validated with 91 clinical samples obtained from patients with malaria compatible symptoms.The technique showed high sensitivity(100%)and specificity(96%)when it was compared to the reference method employed for malaria diagnosis in the Instituto de Salud Carlos栿and a published real-time PCR malaria assay.Conclusions:The technique designed is an economical,sensitive and specific alternative to current diagnosis methods.Furthermore,the method might be tested in knowlesi-malaria endemic areas with a higher number of samples to confirm the quality of the method.

关键词： Malaria Plasmodium knowlesi Nested multiplex PCR Molecular diagnosis

Plant cell wall-mediated disease resistance:Current understanding and future perspectives

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Molecular Plant 2024年第5期17卷 699-724页

作者： Antonio Molina Lucıa Jorda miguel Angel Torres Marina Martın-Dacal Diego Jose Berlanga Patricia Fernandez-Calvo Elena Gomez-rubio Sonsoles Martın-Santamarıa Centro de Biotecnología y Genómica de Plantas Universidad Politécnica de Madrid(UPM)-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria(INIA/CSIC)Campus de Montegancedo UPMPozuelo de Alarcon(Madrid)Spain Departamento de Biotecnologıa-Biología Vegetal Escuela Técnica Superior de Ingenierıa AgronomicaAlimentarıa y de BiosistemasUPMMadridSpain Centro de Investigaciones Biologicas Margarita Salas Consejo Superior de Investigaciones Científicas(CSIC)Ramiro de Maeztu 928040 MadridSpain Misión Biológica de Galicia Consejo Superior de Investigaciones Científicas(CSIC)Campus de Montegancedo UPMPozuelo de Alarcón(Madrid)Spain

Beyond their function as structural barriers,plant cell walls are essential elements for the adaptation of plants to environmental conditions.Cell walls are dynamic structures whose composition and integrity can be altered in response to environmental challenges and developmental cues.These wall changes are perceived by plant sensors/receptors to trigger adaptative responses during development and upon stress perception.Plant cell wall damage caused by pathogen infection,wounding,or other stresses leads to the release of wall molecules,such as carbohydrates(glycans),that function as damage-associated molecular patterns(DAMPs).DAMPs are perceived by the extracellular ectodomains(ECDs)of pattern recognition receptors(PRRs)to activate pattern-triggered immunity(PTI)and disease resistance.Similarly,glycans released from the walls and extracellular layers of microorganisms interacting with plants are recognized as microbe-associated molecular patterns(MAMPs)by specific ECD-PRRs triggering PTI responses.The number of oligosaccharides DAMPs/MAMPs identified that are perceived by plants has increased in recent years.However,the structural mechanisms underlying glycan recognition by plant PRRs remain limited.Currently,this knowledge is mainly focused on receptors of the LysM-PRR family,which are involved in the perception of various molecules,such as chitooligosaccharides from fungi and lipo-chitooligosaccharides(i.e.,Nod/MYC factors from bacteria and mycorrhiza,respectively)that trigger differential physiological responses.Nevertheless,additional families of plant PRRs have recently been implicated in oligosaccharide/polysaccharide recognition.These include receptor kinases(RKs)with leucine-rich repeat and Malectin domains in their ECDs(LRR-MAL RKs),Catharanthus roseus RECEPTOR-LIKE KINASE 1-LIKE group(CrRLK1L)with Malectin-like domains in their ECDs,as well as wall-associated kinases,lectin-RKs,and LRR-extensins.The characterization of structural basis of glycans recognition by these new plant receptors will shed light

关键词： Arabidopsis thaliana cell wall damage-associated molecular patterns DAMPs disease resistance glycans oligosaccharides pattern-triggered immunity PTl pattern recognition receptor PRRs

A prototype for on-site generation of chlorinated disinfectant for use in rural aqueducts

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Water Science and Engineering 2024年第1期17卷 33-40页

作者： Diana Marcela Cuesta Parra Felipe Correa Mahecha Andrés Felipe rubio Pinzon Davidcamilo Ramirez Bustos Leonel Alveyro Teran Llorente miguel Fernando Jimenez Jimenez Chemical and Environmental Engineering Department Fundacion Universidad de AmericaBogota 11711Colombia Chemical Engineering Department Fundacion Universidad de AmericaBogota 117111Colombia Mecanical Engineering Department Fundacion Universidad de AmericaBogota 117111Colombia

Sodium hypochlorite has significant potential as a sanitation solution in hard-to-reach areas.Few studies have investigated the optimal electrolysis parameters for its production with volumes greater than 1o L.This study evaluated sodium hypochlorite production through electrolysis in a 22-L prototype and identified the optimal operating parameters.Tests were performed using graphite electrodes with areas of 68.4 cm²at the laboratory scale and 1865.0 cm?at the prototype scale.A design for experiments with different operating times,chloride concentrations,and electric current intensities was developed.The optimal operating time,sodium chloride concentration,and current intensity at the laboratory scale were 120 min,150 g of chloride per liter,and 3 A,respectively,leading to the production of 5.02 g/L of the disinfectant with an energy efficiency of 12.21 mg of Cl2 per kilojoule.At the prototype scale,the maximum sodium hypochlorite concentration of 3.99 g of chloride per liter was achieved with an operating time of 120 min,a sodium chloride concentration of 100 g of chloride per liter,and a current intensity of 70 A,reaching an energy efficiency of 42.56 mg of Cl2 per kilojoule.In addition,this study evaluated the influences of the chloride concentration,current intensity,and operating time on the production of sodium hypochlorite at the two scales,and formulated the equations showing the trends of sodium hypochlorite production and energy efficiency in the electrochemical systems.The 22-L prototype model for production of this oxidizing substance is promising for disinfection of large volumes of water in areas that are difficult to access.

关键词： Disinfection Electrolysis Energy efficiency Optimization Sodium hypochlorite

Re-folded structure of syn-orogenic granitoids(Padrón dome, NW Iberia):Assessing rheological evolution of cooling continental crust in a collisional setting

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Geoscience Frontiers 2019年第2期10卷 651-669页

作者： Rubén Díez Fernández Francisco J.rubio Pascual Luis miguel Martín Parra Departamento de Geodinámica Estratigrafía y PaleontologíaFacultad de Ciencias GeológicasUniversidad Complutense de Madrid Instituto Geológico y Minero de Espana

This contribution discusses about the rheological, kinematic and dynamic frameworks necessary to produce recumbent and upright folds from syn-orogenic granitic massifs that were formed during an early stage of magma genesis related to the onset of a migmatitic dome. Syn-kinematic granitoids occurring within the high-grade infrastructure of the Padron migmatitic dome(NW Iberia) are deformed into largescale recumbent folds(D_2) that are later affected by upright folds(D_3). Petrostructural analysis of a selected area of this dome reveals that after a period of crustal thickening(D_1), NNW-directed extensional flow gave way to recumbent folds and penetrative axial plane foliation(S_2). Superimposed subhorizontal compression resulted in upright folds(D_3). A closer view into the dynamics of the dome allows exploring the factors that may condition the nucleation of folds with contrasting geometries during progressive deformation of molten continental crust. The formation of folds affecting syn-kinematic granitoids suggests a cooling metamorphic path in migmatitic domes. Active and passive folding mechanisms require a crystallizing(cooling) magma to nucleate folds. A more competent metamorphic host inhibits fold nucleation from much less competent magmas. As it crystallizes, magma becomes more rigid(competent),and approaches viscosity values of its host. Passive folding is favored when no significant competence contrast exists between magma and host, so this folding mechanism is more likely shortly after magma genesis and emplacement. In such conditions, and under dominant subhorizontal flow accompanied by flattening(D_2),passive folding would produce isoclinal recumbent geometries. Further magma cooling introduces a shift into the rheological behavior of partially molten crust. Thereon, crystallizing magma bodies would represent significant competence contrasts relative to their host. At this point, buckling is a more likely folding mechanism, and more regular, buckle folds re-fold previous structures after significan

关键词： Recumbent folds Migmatitic domes Partial melting Crustal-derived granitoids Orogenic collapse Variscan orogen

Role of cetuximab in first-line treatment of metastatic colorectal cancer

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World Journal of Gastroenterology 2014年第15期20卷 4208-4219页

作者： miguel Jhonatan Sotelo Beatriz García-Paredes Carlos Aguado Javier Sastre Eduardo Díaz-rubio Center affiliate to the "Red Temática de Investigación Cooperativa (RD12/0036/006)" Instituto Carlos ⅢSpanish Ministry of Economy and Competitivity. Oncology DepartmentHospital Clínico San Carlos28040 MadridSpain Department of Medicine School of MedicineUniversidad Complutense de Madrid28040 MadridSpain

The treatment of metastatic colorectal cancer(mCRC)has evolved considerably in the last decade,currently allowing most mCRC patients to live more than two years.Monoclonal antibodies targeting the epidermal growth factor receptor(EGFR)and vascular endothelial growth factor play an important role in the current treatment of these patients.However,only antibodies directed against EGFR have a predictive marker of response,which is the mutation status of v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog(KRAS).Cetuximab has been shown to be effective in patients with KRAS wild-type mCRC.The CRYSTAL study showed that adding cetuximab to FOLFIRI(regimen of irinotecan,infusional fluorouracil and leucovorin)significantly improved results in the first-line treatment of KRAS wildtype mCRC.However,results that evaluate the efficacy of cetuximab in combination with oxaliplatin-based chemotherapy in this setting are contradictory.On the other hand,recent advances in the management of colorectal liver metastases have improved survival in these patients.Adding cetuximab to standard chemotherapy increases the response rate in patients with wild-type KRAS and can thus increase the resectability rate of liver metastases in this group of patients.In this paper we review the different studies assessing the efficacy of cetuximab in the first-line treatment of mCRC.

关键词： Cetuximab First-line Metastatic colorectal cancer

Should capecitabine replace 5-fluorouracil in the first-line treatment of metastatic colorectal cancer?

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World Journal of Gastroenterology 2014年第20期20卷 6092-6101页

作者： Carlos Aguado Beatriz García-Paredes miguel Jhonatan Sotelo Javier Sastre Eduardo Díaz-rubio Department of Medical Oncology Hospital Clínico San Carlos 28040 Madrid Spain

Fluoropyrimidines play a central role in the first-line treatment of metastatic colorectal cancer.Our aim was to review whether capecitabine was a safer,non-inferior,economically superior and more convenient alternative to 5-fluorouracil.Capecitabine has previously been compared to 5-fluorouracil-either as a monotherapy or in combination with oxaliplatin,irinotecan,or biological drugs-and has been found to have comparable efficacy and safety profiles.Furthermore,pharmacoeconomic data and patients’preferences for oral chemotherapy further favor capecitabine.Therefore,capecitabine appears to be an effective and safe alternative to fluorouracil in the first-line treatment of metastatic colorectal cancer.

关键词： Capecitabine 5-Fluorouracil Metastatic colorectal

Translational pancreatic cancer research:a comparative study on patient-derived xenograft models

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World Journal of Gastroenterology 2018年第7期24卷 794-809页

作者： Mercedes rubio-Manzanares Dorado Luis miguel Marín Gómez Daniel Aparicio Sánchez Sheila Pereira Arenas Juan Manuel Praena-Fernández Juan Jose Borrero Martín Francisco Farfán López miguel ángel Gómez Bravo Jordi Muntané Relat Javier Padillo Ruiz Department of Hepatobiliary and Pancreatic Surgery Virgen del Rocio University Hospital Oncology Surgery Cell Therapy and Organ Transplantation Group Institute of Biomedicine of Seville (IBiS) Virgen del Rocio University Hospital University of Seville Statistics Methodology and Evaluation of Research Unit Virgen del Rocio University Hospital Pathology Department Virgen del Rocio University Hospital

AIM To assess the viability of orthotopic and heterotopic patient-derived pancreatic cancer xenografts implanted into nude mice.METHODS This study presents a prospective experimental analytical follow-up of the development of tumours in mice upon implantation of human pancreatic adenocarcinoma samples. Specimens were obtained surgically from patients with a pathological diagnosis of pancreatic adenocarcinoma. Tumour samples from pancreatic cancer patients were transplanted into nude mice in three different locations(intraperitoneal, subcutaneous and pancreatic). Histological analysis(haematoxylin-eosin and Masson's trichrome staining) and immunohistochemical assessment of apoptosis(TUNEL), proliferation(Ki-67), angiogenesis(CD31) and fibrogenesis(α-SMA) were performed. When a tumour xenograft reached the target size, it was reimplanted in a new nude mouse. Three sequential tumour xenograft generations were generated(F1, F2 and F3).RESULTS The overall tumour engraftment rate was 61.1%. The subcutaneous model was most effective in terms of tissue growth(69.9%), followed by intraperitoneal(57.6%) and pancreatic(55%) models. Tumour development was faster in the subcutaneous model(17.7 ± 2.6 wk) compared with the pancreatic(23.1 ± 2.3 wk) and intraperitoneal(25.0 ± 2.7 wk) models(P = 0.064). There was a progressive increase in the tumour engraftment rate over successive generations for all three models(F1 28.1% vs F2 71.4% vs F3 80.9%, P < 0.001). There were no significant differences in tumour xenograft differentiation and cell proliferation between human samples and the three experimental models among the sequential generations of tumour xenografts. However, a progressive decrease in fibrosis, fibrogenesis, tumour vascularisation and apoptosis was observed in the three experimental models compared with the human samples. All three pancreatic patient-derived xenograft models presented similar histological and immunohistochemical characteristics.CONCLUSION In our experience, the faster development andgreatest number o

关键词： Immunohistological analysis Pancreatic cancer Patient-derived xenograft Animal model Nude mice

Increased postoperative complications after protective ileostomy closure delay: An institutional study

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World Journal of Gastrointestinal Surgery 2014年第9期6卷 169-174页

作者： Ines rubio-Perez miguel Leon Daniel Pastor Joaquin Diaz Dominguez Ramon Cantero General and Digestive Surgery Department La Paz University Hospital

AIM: To study the morbidity and complications as-sociated to ileostomy reversal in colorectal surgery pa-tients, and if these are related to the time of closure. METHODS: A retrospective analysis of 93 patients, who had undergone elective ileostomy closure between 2009 and 2013 was performed. Demographic, clinical and surgical variables were reviewed for analysis. All complications were recorded, and classified according to the Clavien-Dindo Classification. Statistical univariate and multivariate analysis was performed, setting a P value of 0.05 for significance.RESULTS: The patients had a mean age of 60.3 years, 58% male. The main procedure for ileostomy cre-ation was rectal cancer(56%), and 37% had received preoperative chemo-radiotherapy. The average delay from creation to closure of the ileostomy was 10.3 mo. Postoperative complications occurred in 40% of the pa-tients, with 1% mortality. The most frequent were ileus(13%) and wound infection(13%). Pseudomembra-nous colitis appeared in 4%. Increased postoperative complications were associated with delay in ileostomyclosure(P = 0.041). Male patients had more complica-tions(P = 0.042), mainly wound infections(P = 0.007). Pseudomembranous colitis was also associated with the delay in ileostomy closure(P = 0.003). End-to-end in-testinal anastomosis without resection was significantly associated with postoperative ileus(P = 0.037). CONCLUSION: Although closure of a protective il-eostomy is a fairly common surgical procedure, it has a high rate of complications, and this must be taken into account when the indication is made. The delay in stoma closure can increase the rate of complications in general, and specifically wound infections and colitis.

关键词： closure protective postoperative ileus colorectal stoma anastomosis indication univariate multivariate