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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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Vinyl sulfonyl chemistry-driven unidirectional transport of a macrocycle through a[2]rotaxane

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Organic Chemistry Frontiers 2022年第3期9卷 633-642页

作者： Arthur H.G.David pablo García-Cerezo Araceli G.Campaña Francisco santoyo-González Victor Blanco Departamento de Química Orgánica Facultad de CienciasUnidad de Excelencia de Química Aplicada a Biomedicina y Medioambiente(UEQ)Universidad de Granada(UGR)Avda.Fuente Nueva S/N18071 GranadaSpain

By applying a combination of the coupling-and-decoupling(CAD)chemistry of the vinyl sulfonate group with the click thia-Michael addition to the vinyl sulfone group(MAVs)we performed the irreversible unidirectional transportation of the ring through the linear component in a[2]rotaxane by a chemically and pHdriven flashing energy ratchet mechanism.

关键词： mechanism unidirectional chemistry

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Benign course of residual inflammation at end of treatment of liver transplant recipients after sofosbuvir based therapy

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World Journal of Hepatology 2022年第3期14卷 602-611页

作者： Bahaaeldeen Ismail Karim M Benrajab pablo Bejarano Phillip Ruiz Debbie sears Andreas Tzakis Xaralambos Bobby Zervos Division of Digestive Diseases and Nutrition University of Kentucky College of MedicineLexingtonKY 40536United States Department of Pathology Cleveland Clinic FloridaWestonFL 33331United States Department of Pathology University of Miami Miller School of MedicineMiamiFL 33136United States Department of Liver Transplant Cleveland Clinic FloridaWestonFL 33331United States

BACKGROUND Persistent inflammation on histology after successful hepatitis C(HCV)treatment has been reported.However,data regarding the long-term impact in liver transplant recipients is limited,particularly after using direct-acting antiviral(DAA)therapies.AIM To evaluate the impact of successful treatment with DAAs on histological changes and occult HCV and to describe the clinical course of residual inflammation in liver transplant recipients.METHODs We conducted a case series of 13 chronic HCV infected liver transplant recipients successfully treated with DAAs between December 2013 and May 2014.All patients were treated for 24 wk and had non-detectable serum HCV RNA by the time of biopsy.Only patients with at least one liver biopsy at or after treatment were included.We examined liver biopsies for evidence of residual inflammation and the presence of intrahepatic HCV RNA.REsULTs Persistent inflammation was seen in 12/13 patients on end of treatment biopsy.Inflammation was still seen in the available five follow-up biopsies(range 38-48 wk after the end of treatment).Intrahepatic HCV RNA was undetectable in all biopsies.All patients had preserved graft function for a mean follow-up of 2.5 years,except one that developed chronic rejection.CONCLUsION After successful HCV treatment with DAAs,liver transplant recipients may have persistent inflammation on biopsy without evidence of intracellular RNA.The clinical outcome remained favorable in most patients.Further studies with a larger number and longer follow-up are needed to establish the implication of this finding on long-term graft function.

关键词： Immunosuppression Liver transplantation Recurrent hepatitis C sustained virologic response Interferon

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Chemokines and Chemokine Receptors:Their Manifold Roles in Homeostasis and Disease

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Cellular & Molecular Immunology 2004年第2期1卷 95-104页

作者： pablo Iribarren Laboratory of Molecular Immunoregulation Center for Cancer ResearchNational Cancer Institute at FrederickFrederickMarylandUSA

Chemokines are a superfamily of small proteins that bind to G protein-coupled receptors on target cells and were originally discovered as mediators of directional migration of immune cells to sites of inflammation and injury.In recent years,it has become clear that the function of chemokines extends well beyond the role in leukocyte chemotaxis.They participate in organ development,angiogenesis/angiostasis,leukocyte trafficking and homing,tumorigenesis and metastasis,as well as in immune responses to microbial infection.Therefore, chemokines and their receptors are important targets for modulation of host responses in pathophysiologicai conditions and for therapeutic intervention of human diseases.Cellular & Molecular Immunology.2004;1(2): 95-104.

关键词： chemokine receptor development leukocyte homing angiogenesis metastasis

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The glutamate metabotropic receptor 5(GRM5)gene is associated with beef cattle home range and movement tortuosity

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Journal of Animal science and Biotechnology 2023年第1期14卷 72-87页

作者： Cristian A.Moreno Garcia Huitong Zhou David Altimira Robyn Dynes pablo Gregorini sadeepa Jayathunga Thomas M.R.Maxwell Jonathan Hickford Department of Agricultural sciences Faculty of Agriculture and Life SciencesLincoln UniversityPO Box 85084Lincoln 7647New Zealand. Exertion Games Lab 8 Scenic BoulevardClaytonVictoria 3800Australia. AgResearch Limited Lincoln Research CentreCnr Springs Road and Gerald StreetLincoln 7674New Zealand. scion Tītokorangi Drive(formerly Long Mile Rd)Rotorua 3010New Zealand.

Background:The grazing behaviour of herbivores and their grazing personalities might in part be determined genetically,but there are few studies in beef cattle illustrating this.In this study,we investigated for first time the genetic variation within a candidate‘grazing gene',the glutamate metabotropic receptor 5 gene(GRM5),and tested associations between variation in that gene and variation in grazing personality behaviours(GP-behaviours)displayed by free-ranging cows during winter grazing in the steep and rugged rangelands of New Zealand.Mature beef cows(n=303,from 3 to 10 years of age)were tracked with global positioning system(GPs)and,with 5-minutes(min)relocation frequency,various GP-behaviours were calculated.These included horizontal and vertical distances travelled,mean elevation,elevation range,elevation gain,slope,home range and movement tortuosity,variously calculated using daily relocation trajectories with repeated measurements(i.e.,7 to 24 days(d))and satellite-derived digital elevation models(DEM).The different GP-behaviours were fitted into mixed models to ascertain their associations with variant sequences and genotypes of GRM5.Results:We discovered three GRM5 variants(A,B and C)and identified the six possible genotypes in the cattle studied.The mixed models revealed that A was significantly associated with elevation range,home range and movement tortuosity.similarly,GRM5 genotypes were associated(Psity,while trends suggesting association(Pso revealed for elevation range and horizontal distance travelled.Most GP-behaviour models were improved by correcting for cow age-class as a fixed factor.The analysis of GP-behaviours averaged per cow age-class suggests that grazing personality is fully established as beef cows reached 4 years of age.Home range and movement tortuosity were not only associated with GRM5 variation,but also negatively correlated with each other(r=-0.27,Psions:There seems to be a genetically determined trade-off bet

关键词： Animal personality Breeding programmes Genetic associations Grazing distribution Grazing patterns steep and rugged terrain

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HVOF-sprayed HAp/s53P4 BG composite coatings on an AZ31 alloy for potential applications in temporary implants

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Journal of Magnesium and Alloys 2024年第1期12卷 345-360页

作者： Carlos A.Poblano-salas John Henao Astrid L.Giraldo-Betancur Paola Forero-sossa Diego German Espinosa-Arbelaez Jorge A.González-sánchez Luis R.Dzib-Pérez susana T.Estrada-Moo Idelfonso E.Pech-Pech CIATEQ A.C. Av.Manantiales 23-AParque Industrial Bernardo QuintanaEl MarquésQuerétaro 76246Mexico CONAHCYT-CIATEQ A.C. Av.Manantiales 23-AParque Industrial Bernardo QuintanaEl MarquésQuerétaro 76246Mexico CONAHCYT-Cinvestav Libramiento Norponiente#2000.Fraccionamiento Real de JuriquillaQuerétaro 76230Mexico Universidad Tecnológica de Querétaro Av.Pie de la Cuesta N°2501Querétaro 76148Mexico CIDEsI Av.Playa Pie de la Cuesta No 702Col.San PabloQuerétaro 76125Mexico Center for Corrosion Research Autonomous University of CampecheAv.Héroe de Nacozari No.480P.C.24070San Francisco de CampecheCam.Mexico

Bioactive thermal spray coatings produced via high-velocity oxygen fuel spray(HVOF)from hydroxyapatite(HAp)and bioactive glasses(BG)have the potential to be employed on temporary implants due to the ability of both HAp and BG to dissolve and promote osseointegration,considering that both phases have different reaction and dissolution rates under in-vitro conditions.In the present work,75%wt.HAp-25%wt.s53P4 bioactive glass powders were HVOF-sprayed to obtain HAp/s53P4 BG composite coatings on a bioresorbable AZ31 alloy.The study is focused on exploring the effect of the stand-off distance and fuel/oxygen ratio variation as HVOF parameters to obtain stable structural coatings and to establish their effect on the phases and microstructure produced in those coatings.Different characterization techniques,such as scanning electron microscopy,X-ray diffraction,and Fourier transform infrared spectroscopy,were employed to characterize relevant structural and microstructural properties of the composite coatings.The results showed that thermal gradients during coating deposition must be managed to avoid delamination due to the high temperature achieved(max 550℃)and the differences in coefficients of thermal expansion.It was also found that both spraying distance and oxygen/fuel ratio allowed to keep the hydroxyapatite as the main phase in the coatings.In addition,in-vitro electrochemical studies were performed on the obtained HAp/s53P4 BG composite coatings and compared against the uncoated AZ31 alloy.The results showed a significant decrease in hydrogen evolution(at least 98%)when the bioactive coating was applied on the Mg alloy during evaluation in simulated body fluid(sBF).

关键词： Coatings Composites Thermal spray Temporary implants Hydrogen evolution

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Asymptotic of Eigenvalues and Lattice Points

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Acta Mathematica sinica,English series 2006年第6期22卷 1645-1650页

作者： Juan pablo PINAsCO Instituto de Ciencias Universidad Nacional de General Sarmiento

dimension. elementary domains of In this work we study the spectral counting function for the p-Laplace operator in one We show the existence of a two-term Weyl-type asymptote. The method of proof is rather based on ... 详细信息

关键词： p-Laplacian eigenvalues lattice points

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Parallel array with axially coded light-sheet microscope

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Light(science & Applications) 2020年第1期9卷 1387-1388页

作者： pablo Loza-Alvarez ICFO-The Institute of Photonic sciences The Barcelona Institute of Science and Technology08860 CastelldefelsBarcelonaSpain

A parallel array of frequency modulated light sheets results in a scanning-less light sheet microscope capable of fast volumetric imaging.

关键词： array light axial

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Parkinson’s disease and diabetes mellitus: common mechanisms and treatment repurposing

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Neural Regeneration Research 2022年第8期17卷 1652-1658页

作者： Carmen M.Labandeira Arturo Fraga-Bau David Arias Ron Elena Alvarez-Rodriguez pablo Vicente-Alba Javier Lago-Garma Ana I.Rodriguez-Perez Department of Clinical Neurology Hospital Alvaro CunqueiroUniversity Hospital ComplexVigoSpain Laboratory of Cellular and Molecular Neurobiology of Parkinson’s Disease Research Center for Molecular Medicine and Chronic Diseases(CIMUS)IDISUniversity of Santiago de CompostelaSantiago de CompostelaSpain Department of Clinical Oncology University Hospital ComplexOurenseSpain Department of Endocrinology Hospital MeixoeiroUniversity Hospital ComplexVigoSpain Networking Research Center on Neurodegenerative Diseases(CiberNed) MadridSpain

In the last decade,attention has become greater to the relationship between neurodegeneration and abnormal insulin signaling in the central nervous system,as insulin in the brain is implicated in neuronal survival,plasticity,oxidative stress and neuroinflammation.Diabetes mellitus and Parkinson’s disease are both aging-associated diseases that are turning into epidemics worldwide.Diabetes mellitus and insulin resistance not only increase the possibility of developing Parkinson’s disease but can also determine the prognosis and progression of Parkinsonian symptoms.Today,there are no available curative or disease modifying treatments for Parkinson’s disease,but the role of insulin and antidiabetic medications in neurodegeneration opens a door to treatment repurposing to fight against Parkinson’s disease,both in diabetic and nondiabetic Parkinsonian patients.Furthermore,it is essential to comprehend how a frequent and treatable disease such as diabetes can influence the progression of neurodegeneration in a challenging disease such as Parkinson’s disease.Here,we review the present evidence on the connection between Parkinson’s disease and diabetes and the consequential implications of the existing antidiabetic molecules in the severity and development of Parkinsonism,with a particular focus on glucagon-like peptide-1 receptor agonists.

关键词： antidiabetic diabetes mellitus dopamine exenatide glucagon-like peptide-1 insulin neurodegeneration neuroinflammation Parkinson’s disease repurposing

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Upper gastrointestinal endoscopic findings in celiac disease at diagnosis:A multicenter international retrospective study

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World Journal of Gastroenterology 2022年第43期28卷 6157-6167页

作者： Juan pablo stefanolo Fabiana Zingone Carolina Gizzi Ilaria Marsilio María Luján Espinet Edgardo Gustavosmecuol Mark Khaouli María Laura Moreno María I Pinto-sánchez sonia Isabel Niveloni Elena F Verdú Carolina Ciacci Julio César Bai small Bowel section Dr.C.Bonorino Udaondo Gastroenterology HospitalBuenos Aires 1264Argentina Department of surgery Oncology and GastroenterologyUniversity of PaduaPadova 35124Italy Gastroenterology Unit Azienda Ospedale UniversitàPadova 35128Italy Department of Medicine SurgeryDentistryScuola Medica SalernitanaUniversity of SalernoSalerno 84081Italy Department of Medicine Farncombe Family Digestive Health Research InstituteMcMaster UniversityHamilton L8S 4K1Canada Research Institutes Universidad del SalvadorBuenos Aires 1020Argentina

BACKGROUND Gastroduodenal endoscopy and biopsy following positive specific serology is considered the gold standard to diagnose celiac disease(CeD)in adults.Whether upper endoscopy helps detect comorbid conditions is unknown.AIM To investigate the prevalence of non-celiac endoscopic findings in patients in whom endoscopy was performed to confirm CeD diagnosis.METHODs This is an observational,descriptive,multicenter,retrospective study that reports endoscopic findings obtained in adult patients enrolled in local registries from four tertiary centers.We collected data reported on first endoscopy,indicated for investigation of CeD.Diagnosis of CeD was performed by histology(≥Marsh 2type mucosal damage)and specific serology.Two European and one North American center included biopsy-confirmed CeD following positive serology.A fourth center(south America)included symptomatic patients undergoing endoscopy,irrespective of CeD serology.The latter cohort included a non-CeD control group.REsULTs A total of 1328 patients(80%female;35 years median age)were enrolled,of whom 95.6%had positive specific serology.In 135 patients,endoscopy revealed 163 abnormalities unrelated to CeD(prevalence:10.1%).Erosive reflux esophagitis(6.4%),gastric erosions(2.0%),and suspicion of esophageal metaplasia(1.2%)were the most common findings.Biopsy-confirmed Barrett’s esophagus was infrequent(0.2%).No endoscopic cancer was detected.Older patients(≥51 years of age)had a higher prevalence of endoscopic findings than those≤50(Psouth American cohort,CeD was associated with a lower rate(8.2%)of comorbid endoscopic findings compared with controls(29.1%;Psted multivariate analysis of this cohort,having CeD was associated with a 72%reduction in the risk of any endoscopic abnormality(Psymptoms was associated with a 37%reduction in the risk of finding at least one endoscopic lesion(PsION In this large multicenter study,young adults with positive CeD serology had few comorbid endos

关键词： Celiac disease Upper gastrointestinal endoscopy Concomitant endoscopic lesions Malignancies Multicenter study

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