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CRISPR/Cas9 editing of the polygalacturonase FaPG1 gene improves strawberry fruit firmness

Horticulture Research 2023年第3期10卷 201-210页

作者： Gloria López-Casado Cristina Sánchez-Raya pablo d.ric-varas Candelas Paniagua Rosario Blanco-Portales Juan Muñoz-Blanco Sara Pose Antonio J.Matas Jose A.Mercado departamento de Botánica y Fisiología Vegetal Instituto de Hortofruticultura Subtropical y Mediterránea‘La Mayora’(IHSM-UMA-CSIC)Universidad de Málaga29071 MálagaSpain departamento de Bioquímica y Biología Molecular Universidad de Córdoba14071CórdobaSpain

Firmness is one of the most important fruit quality traits in strawberries.The postharvest shelf life of this soft fruit is highly limited by the loss of firmness,where cell wall disassembly plays an important role.Previous studies demonstrated that the polygalacturonase FaPG1 has a key role in remodelling pectins during strawberry softening.In this study,FaPG1 knockout strawberry plants have been generated using the CRISPR/Cas9 system delivered via Agrobacterium tumefaciens.Ten independent lines,cv.“Chandler”,were obtained,and all of them were successfully edited as determined by PCR amplification and T7 endonuclease assay.The targeted mutagenesis insertion and deletion rates were analyzed using targeted deep sequencing.The percentage of edited sequences varied from 47%up to almost 100%,being higher than 95%for seven of the selected lines.Phenotypic analyses showed that 7 out of the eight lines analyzed produced fruits significantly firmer than the control,ranging from 33 to 70%increase in firmness.There was a positive relationship between the degree of FaPG1 editing and the rise in fruit firmness.Minor changes were observed in other fruit quality traits,such as colour,soluble solids,titratable acidity or anthocyanin content.Edited fruits showed a reduced softening rate during postharvest,displayed a reduced transpirational water loss,and were less damaged by Botrytis cinerea inoculation.The analysis of four potential off-target sites revealed no mutation events.In conclusion,editing the FaPG1 gene using the CRISPR/Cas9 system is an efficient method for improving strawberry fruit firmness and shelf life.

关键词： fir straw PG1

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Blockade of Rho-associated kinase prevents inhibition of axon regeneration of peripheral nerves induced by anti-ganglioside antibodies

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Neural Regeneration Research 2024年第4期19卷 895-899页

作者： Andrés Berardo Cristian R.Bacaglio Bárbara B.Báez Rubén Sambuelli Kazim A.Sheikh pablo H.H.Lopez Laboratorio de Neurobiología Instituto de Investigación Médica Mercedes y Martin Ferreyra(INIMEC)-Consejo Nacional de Investigaciones Cientificas y Técnicas(CONICET)Universidad Nacional de Córdoba departamento de Química Biológica-Centro de Investigaciones en Química Biológica de Córdoba(CIQUIBIC)-Consejo Nacional de Investigaciones Cientificas y Técnicas(CONICET) Facultad de Cs.QuímicasUniversidad Nacional de Córdoba Servicio de Anatomía Patológica Clínica Universitaria Reina FabiolaUniversidad Católica de Córdoba department of Neurology University of Texas Medical School at Houston

Anti-ganglioside antibodies are associated with delayed/poor clinical recovery in Guillain-Barrèsyndrome,mostly related to halted axon regeneration.Cross-linking of cell surface gangliosides by anti-ganglioside antibodies triggers inhibition of nerve repair in in vitro and in vivo paradigms of axon regeneration.These effects involve the activation of the small GTPase Rho A/ROCK signaling pathways,which negatively modulate growth cone cytoskeleton,similarly to well stablished inhibitors of axon regeneration described so far.The aim of this work was to perform a proof of concept study to demonstrate the effectiveness of Y-27632,a selective pharmacological inhibitor of ROCK,in a mouse model of axon regeneration of peripheral nerves,where the passive immunization with a monoclonal antibody targeting gangliosides Gd1a and GT1b was previously reported to exert a potent inhibitory effect on regeneration of both myelinated and unmyelinated fibers.Our results demonstrate a differential sensitivity of myelinated and unmyelinated axons to the pro-regenerative effect of Y-27632.Treatment with a total dosage of 9 mg/kg of Y-27632 resulted in a complete prevention of anti-Gd1a/GT1b monoclonal antibody-mediated inhibition of axon regeneration of unmyelinated fibers to skin and the functional recovery of mechanical cutaneous sensitivity.In contrast,the same dose showed toxic effects on the regeneration of myelinated fibers.Interestingly,scale down of the dosage of Y-27632 to 5 mg/kg resulted in a significant although not complete recovery of regenerated myelinated axons exposed to anti-Gd1a/GT1b monoclonal antibody in the absence of toxicity in animals exposed to only Y-27632.Overall,these findings confirm the in vivo participation of Rho A/ROCK signaling pathways in the molecular mechanisms associated with the inhibition of axon regeneration induced by anti-Gd1a/GT1b monoclonal antibody.Our findings open the possibility of therapeutic pharmacological intervention targeting Rho A/Rock pathway in immune neuropathies associated wi

关键词： anti-ganglioside antibodies anti-glycan antibodies axon regeneration ganglioside Guillain-Barrésyndrome nerve repair ROCK Y-27632

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Transarterial embolization is an acceptable bridging therapy to hepatocellular carcinoma prior to liver transplantation

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World Journal of Transplantation 2024年第2期14卷 126-134页

作者： Gabriel Lazzarotto-da-Silva Leandro A Scaffaro Mauricio Farenzena Lucas Prediger Rafaela K Silva Flávia Heinz Feier Tomaz J M Grezzana-Filho pablo d Rodrigues Alexandre de Araujo Mario Reis Alvares-da-Silva Roberta C Marchiori Cleber Rosito Pinto Kruel Marcio Fernandes Chedid department of Liver Transplant and Hepatobiliary Surgery Unit Hospital de Clínicas de Porto AlegrePorto Alegre 90035-007Brazil department of Interventional Radiology Unit Hospital de Clínicas de Porto AlegrePorto Alegre 90035-007Brazil department of Gastroenterology and Hepatology Hospital de Clínicas de Porto AlegrePorto Alegre 90035-007Brazil

BACKGROUNd Hepatocellular carcinoma(HCC)is an aggressive malignant neoplasm that requires liver transplantation(LT).despite patients with HCC being prioritized by most organ allocation systems worldwide,they still have to wait for long periods.Locoregional therapies(LRTs)are employed as bridging therapies in patients with HCC awaiting LT.Although largely used in the past,transarterial embolization(TAE)has been replaced by transarterial chemoembolization(TACE).However,the superiority of TACE over TAE has not been consistently shown in the literature.AIM To compare the outcomes of TACE and TAE in patients with HCC awaiting LT.METHOdS All consecutive patients with HCC awaiting LT between 2011 and 2020 at a single center were included.All patients underwent LRT with either TACE or TAE.Some patients also underwent percutaneous ethanol injection(PEI),concom-itantly or in different treatment sessions.The choice of LRT for each HCC nodule was determined by a multidisciplinary consensus.The primary outcome was waitlist dropout due to tumor progression,and the secondary outcome was the occurrence of adverse events.In the subset of patients who underwent LT,complete pathological response and post-transplant recurrence-free survival were also assessed.RESULTS Twelve(18.5%)patients in the TACE group(only TACE and TACE+PEI;n=65)and 3(7.9%)patients in the TAE group(only TAE and TAE+PEI;n=38)dropped out of the waitlist due to tumor progression(P log-rank test=0.29).Adverse events occurred in 8(12.3%)and 2(5.3%)patients in the TACE and TAE groups,respectively(P=0.316).Forty-eight(73.8%)of the 65 patients in the TACE group and 29(76.3%)of the 38 patients in the TAE group underwent LT(P=0.818).Among these patients,complete pathological response was detected in 7(14.6%)and 9(31%)patients in the TACE and TAE groups,respectively(P=0.145).Post-LT,HCC recurred in 9(18.8%)and 4(13.8%)patients in the TACE and TAE groups,respectively(P=0.756).Posttransplant recurrence-free survival was similar between the groups(P log-rank test=0.71).CONCL

关键词： Hepatocellular carcinoma Transarterial embolization Transarterial chemoembolization Liver transplantation Locoregional therapy Bridging

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Hepatocellular carcinoma in non-alcoholic fatty liver disease from a clinical and public health perspective

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Hepatobiliary Surgery and Nutrition 2022年第5期11卷 755-758页

作者： Luis Antonio díaz Marco Arrese Juan pablo Arab departamento de Gastroenterología Escuela de MedicinaPontificia Universidad Católica de ChileSantiagoChile division of Gastroenterology Department of MedicineSchulich School of MedicineWestern University&London Health Sciences CentreLondonONCanada Alimentiv LondonONCanada

Non-alcoholic fatty liver disease(NAFLd)is the leading cause of chronic liver disease in the western world(1).Currently,the global prevalence of NAFLd is estimated at 30%and is notably higher in South America and the Middle East(1).NAFLd prevalence has been rising during the last decades,and cases of decompensated NAFLd cirrhosis could increase by 168%in 2030(Figure 1A)(2).These trends are probably explained by several factors including the decrease in viral hepatitis incidence and the increasing prevalence of obesity and type 2 diabetes mellitus(T2dM)among the general population(2).

关键词： prevalence NAFLd liver

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Influence of Angiotensin II on α₁-Adrenergic Receptors Function in Rat Aorta and Expression in Vascular Smooth Muscle Cells

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Journal of Biosciences and Medicines 2024年第4期12卷 123-134页

作者： Itzell Alejandrina Gallardo-Ortíz Juan pablo de Jesús Benítez-Garrido Santiago C. Sigrist-Flores Juan Javier López-Guerrero Enrique Hong Rafael Villalobos-Molina Unidad de Biomedicina Facultad de Estudios Superiores Iztacala Universidad Nacional Aut&#243noma de M&#233xico Tlalnepantla M&#233xico Carrera de Enfermerí a Facultad de Estudios Superiores Iztacala Universidad Nacional Aut&#243noma de M&#233xico Tlalnepantla M&#233xico departamento de Farmacobiologí a Centro de Investigaci&#243n and de Estudios Avanzados-IPN Ciudad de M&#233xico M&#233xico Carrera de Mé dico Cirujano Facultad de Estudios Superiores Iztacala Universidad Nacional Aut&#243noma de M&#233xico Tlalnepantla M&#233xico

Angiotensin II (Ang II) is the main mediator of the Renin-Angiotensin-System acting on AT1 and other AT receptors. It is regarded as a pleiotropic agent that induces many actions, including functioning as a growth factor, and as a contractile hormone, among others. The aim of this work was to examine the impact of Ang II on the expression and function of α1-adrenergic receptors (α1-ARs) in cultured rat aorta, and aorta-derived smooth muscle cells. Isolated Wistar rat aorta was incubated for 24 h in dMEM at 37˚C, then subjected to isometric tension and to the action of added norepinephrine, in concentration-response curves. Ang II was added (1 × 10−5 M), and in some experiments, 5-Methylurapidil (α1A-AR antagonist), AH11110A (α1B-AR antagonist), or BMY-7378 (α1d-AR antagonist), were used to identify the α1-AR involved in the response. desensitization of the contractile response to norepinephrine was observed due to incubation time, and by the Ang II action. α1d-AR was protected from desensitization by BMY-7378;while RS-100329 and prazosin partially mitigated desensitization. In another set of experiments, isolated aorta-derived smooth muscle cells were exposed to Ang II and α1-ARs proteins were evaluated. α1d-AR increased at 30 and 60 min post Ang II exposure, the α1A-AR diminished from 1 to 4 h, while α1B-AR remained unchanged over 24 h of Ang II exposure. Ang II induced an increase of α1d-AR at short times, and BMY-7378 protected α1d-AR from desensitization.

关键词： Angiotensin II α1d-AR α1-AR Expression Rat aorta Smooth Muscle Cells

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South Asian dairy smallholders: A scoping review of practices and zoonoses

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Asian Pacific Journal of Tropical Medicine 2023年第10期16卷 446-452页

作者： Emily Schembri Angus J d Campbell Juan pablo Villanueva-Cabezas Melbourne School of Population and Global Health The University of MelbourneVictoriaAustralia The Nossal Institute for Global Health The University of MelbourneVictoriaAustralia The Peter doherty Institute for Infection and Immunity The University of Melbourne and the Royal Melbourne HospitalVictoriaAustralia

Objective: To identify and discuss on-farm management practices linked to bacterial zoonosis risk in smallholder dairy farmers in South Asia. Methods: This scoping review was conducted as per the PRISMA-ScR guidelines. Five hundred and two publications were retrieved from five online databases using a comprehensive search strategy. Studies were selected if they discussed a farm management practice which impacted human health within a South Asian country. Results: Twenty-two studies were included. Seven management practices relevant to farmers, livestock and their shared environment were identified including raw milk consumption, farm hygiene management, personal protective equipment uses, animal vaccination, cleaning udders, hand washing and disposal of afterbirth materials. Preventive practices were found to be utilized at lower frequencies compared to risk increasing practices. Awareness of bacterial zoonoses is particularly low within the region. Conclusions: Based on the results of this review, it was determined that improving farmer awareness of bacterial zoonotic diseases may favor several of the presented leverage points within the South Asian smallholder dairy system. Relying on formal school education to improve this awareness may not solve this problem, instead, more focus on accessible and affordable zoonoses education and farming programs is required.

关键词： Management practices Zoonoses dairy Smallholder system

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Recent advances in graphene and other 2d materials

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Nano Materials Science 2022年第1期4卷 3-9页

作者： pablo Ares Kostya S.Novoselov department of Physics&Astronomy and National Graphene Institute University of ManchesterManchesterM139PLUK Centre for Advanced 2d Materials National University of Singapore117546Singapore Chongqing 2d Materials Institute Lingjiang New Area Chongqing 400714China

The isolation of the first two-dimensional material, graphene-a monolayer of carbon atoms arranged in a hexagonal lattice-opened new exciting opportunities in the field of condensed matter physics and materials. Its isolation and subsequent studies demonstrated that it was possible to obtain sheets of atomically thin crystals and that these were stable, and they also began to show its outstanding properties, thus opening the door to a whole new family of materials, known as two-dimensional materials or 2d materials. The great interest in different 2d materials is motivated by the variety of properties they show, being candidates for numerous applications.Additionally, the combination of 2d crystals allows the assembly of composite, on-demand materials, known as van der Waals heterostructures, which take advantage of the properties of those materials to create functionalities that otherwise would not be accessible. For example, the combination of 2d materials, which can be done with high precision, is opening up opportunities for the study of new challenges in fundamental physics and novel applications. Here we review the latest fundamental discoveries in the area of 2d materials and offer a perspective on the future of the field.

关键词： 2d materials Graphene Molybdenum disulphide Tarition mebl dichalogenides Hexagonal boron nitride van der waals heterostructus Ulrathin nanomaterials

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Controlling flowering of Medicago sativa(alfalfa)by inducing dominant mutations

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Journal of Integrative Plant Biology 2022年第2期64卷 205-214页

作者： Maurizio Junior Chiurazzi Anton Frisgaard Nørrevang Pedro García pablo d.Cerdán Michael Palmgren Stephan Wenkel NovoCrops Center University of CopenhagenThorvaldsensvej 40Frederiksberg C1871 Denmark Copenhagen Plant Science Centre University of CopenhagenThorvaldsensvej 40Frederiksberg C1871 Denmark department of Plant and Environmental Sciences University of CopenhagenThorvaldsensvej 40Frederiksberg C1871 Denmark Fundación Instituto Leloir IIBBA-CONICETAvenida Patricias Argentinas 435Buenos Aires1405 Argentina

Breeding plants with polyploid genomes is challenging because functional redundancy hampers the identification of loss-of-function mutants.Medicago sativa is tetraploid and obligate outcrossing,which together with inbreeding depression complicates traditional breeding approaches in obtaining plants with a stable growth habit.Inducing dominant mutations would provide an alternative strategy to introduce domestication traits in plants with high gene redundancy.Here we describe two complementary strategies to induce dominant mutations in the M.sativa genome and how they can be relevant in the control of flowering time.First,we outline a genome-engineering strategy that harnesses the use of microProteins as developmental regulators.MicroProteins are small proteins that appeared during genome evolution from genes encoding larger proteins.Genomeengineering allows us to retrace evolution and create microProtein-coding genes de novo.Second,we provide an inventory of genes regulated by microRNAs that control plant development.Making respective gene transcripts microRNA-resistant by inducing point mutations can uncouple microRNA regulation.Finally,we investigated the recently published genomes of M.sativa and provide an inventory of breeding targets,some of which,when mutated,are likely to result in dominant traits.

关键词： flowering time genome-engineering Medicago sativa microProtein microRNA

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Marriage Market with Indifferences: A Linear Programming Approach

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Journal of the Operations Research Society of China 2023年第1期11卷 219-242页

作者： Noelia Juarez pablo A.Neme Jorge Oviedo Instituto de Matemática Aplicada San Luis IMASLRedNIEUniversidad Nacional de San Luis and CONICETD5700HHWSan LuisRepublica Argentina

We study stable and strongly stable matchings in the marriage market with indifference in their preferences.We characterize the stable matchings as integer extreme points of a convex polytope.We give an alternative proof for the integrity of the strongly stable matching polytope.Also,we compute men-optimal(women-optimal)stable and strongly stable matchings using linear programming.When preferences are strict,we find the men-optimal(women-optimal)stable matching.

关键词： Matching markets The marriage market with indifferences Optimal stable matchings Linear programming

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Uncovering current pyroregions in Italy using wildfire metrics

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Ecological Processes 2022年第1期11卷 259-275页

作者： Mario Elia Vincenzo Giannico davide Ascoli Juan pablo Argañaraz Marina d’Este Giuseppina Spano Raffaele Lafortezza Giovanni Sanesi department of Agricultural and Environmental Sciences University of Bari A.MoroVia Amendola 165/A70126 BariItaly department of Agricultural Forest and Food SciencesUniversity of TurinLargo Braccini 210095 GrugliascoTOItaly Mario Gulich Institute for Advanced Space Studies(CONAE-UNC) CONICETRoute C45km 8CP 5187 Falda del CañeteCórdobaArgentina department of Geography The University of Hong KongCentennial CampusPokfulam RoadHong KongChina

Background:Pyrogeography is a major field of investigation in wildfire science because of its capacity to describe the spatial and temporal variations of fire disturbance.We propose a systematic pyrogeographic analytical approach to cluster regions on the basis of their pyrosimilarities.We employed the Affinity Propagation algorithm to cluster pyroregions using Italian landscape as a test bed and its current wildfire metrics in terms of density,seasonality and stand replacing fire ratio.A discussion follows on how pyrogeography varies according to differences in the human,biophysical,socioeconomic,and climatic spheres.Results:The algorithm identified seven different pyroregion clusters.Two main gradients were identified that partly explain the variability of wildfire metrics observed in the current pyroregions.First,a gradient characterized by increasing temperatures and exposure to droughts,which coincides with a decreasing latitude,and second,a human pressure gradient displaying increasing population density in areas at lower elevation.These drivers exerted a major influence on wildfire density,burnt area over available fuels and stand replacing,which were associated to warmdry climate and high human pressure.The study statistically highlighted the importance of a North–South gradient,which represents one of the most important drivers of wildfire regimes resulting from the variations in climatic conditions but showing collinearity with socioeconomic aspects as well.Conclusion:Our fully replicable analytical approach can be applied at multiple scales and used for the entire European continent to uncover new and larger pyroregions.This could create a basis for the European Commission to promote innovative and collaborative funding programs between regions that demonstrate pyrosimilarities.

关键词： Pyrogeography Affinity propagation Forest Mediterranean basin Clustering

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