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

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Neural Regeneration Research 2025年第11期20卷 3317-3329页

作者： Jennifer M.Colón-mercado Aranza I.Torrado-Tapias Iris K.Salgado jose M.Santiago Samuel E.Ocasio Rivera Dina P.Bracho-Rincon Luis H.Pagan Rivera Jorge D.Miranda Department of Physiology University of Puerto RicoMedical Sciences CampusSan JuanPRUSA National Institutes of Health National Human Genome Research InstituteBethesdaMDUSA Universidad Central del Caribe School of MedicineBayamónPRUSA University of Puerto Rico Carolina CampusCarolinaPRUSA Institute of Neurobiology San JuanPRUSA

In addition to the loss of motor function,~60% of patients develop pain after spinal cord *** cellular-molecular mechanisms are not well understood,but the data suggests that plasticity within the rostral,epicenter,and caudal penumbra of the injury site initiates a cellularmolecular interplay that acts as a rewiring mechanism leading to central neuropathic *** can lead to the formation of new connections triggering abnormal sensory *** excitatory glutamate transporters are responsible for the reuptake of extracellular glutamate which makes them a critical target to prevent neuronal hyperexcitability and *** previous studies showed a sexually dimorphic therapeutic window for spinal cord injury after treatment with the selective estrogen receptor modulator *** this study,we investigated the anti-allodynic effects of tamoxifen in male and female rats with spinal cord *** hypothesized that tamoxifen exerts anti-allodynic effects by increasing the expression of glutamate transporters,leading to reduced hyperexcitability of the secondary neuron or by decreasing aberrant *** and female rats received a moderate contusion to the thoracic spinal cord followed by subcutaneous slow-release treatment of tamoxifen or matrix pellets as a control(placebo).We used von Frey monofilaments and the“up-down method”to evaluate mechanical *** treatment decreased allodynia only in female rats with spinal cord injury revealing a sexdependent *** expression profile of glutamatergic transporters(excitatory amino acid transporter 1/glutamate aspartate transporter and excitatory amino acid transporter 2/glutamate transporter-1)revealed a sexual dimorphism in the rostral,epicenter,and caudal areas of the spinal cord with a pattern of expression primarily on *** rodents showed a significantly higher level of excitatory amino acid transporter-1 expression while male rodents showed increased

关键词： allodynia central neuropathic pain EAAT-1/GLAST EAAT-2/GLT-1 glutamate transporters selective estrogen receptor modulator sexual dimorphism spinal cord injury trauma

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Theoretical and Experimental Analysis of Vortex Origin

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Journal of Mathematics and System Science 2013年第5期3卷 251-261页

作者： Pedro Guido Aldana jose Roberto mercado Mexican Institute of Water Technology Paseo Cuauhn6huac 8532 Progreso Jiutepec Morelos CP 62550 Mexico

Several investigations refer to the issue of creation and identification of vortices in flows with different regime and presence of obstacles. Reasons have to do with the crucial role that vortices play in nature and industrial processes （sediment transport, mixing, radiation, noise, etc.）. Despite the contributions, further work is needed in order to perform more analysis of the mathematical arguments used to explain this phenomenon. In this idea order, the paper presents some advances in mathematical analysis and experimental results. In the first section, we do a description of the fluid motion from a fractional view through a sequence of three steps： Darcy＇s law, Navier-Stokes equations and Reynolds equations. Next, a representation of the temporal change of kinetic energy is found, which allows the possibility of the two signs. We obtain a description of the process of vortex creation. A length that represents the transition between flow and vortex intensity is found; then a succession of lengths is established that allows scaling from micro to macro. In the second section, experimental results are present; we consider vortex creation and its detection upstream of a bed form similar to that found in rivers, installed in an open channel, equipped with a water circulation system. For vortex detection, a methodology based on the particle image velocimetry PIV technique is proposed. So, we fulfill two objectives： vortex identification and its passage frequencies behind the bed form installed in the channel. Such procedure allows a computer process time reduction in vortices identification task.

关键词： Navier-Stokes fractional equation Darcy's law vortex detection particle image velocimetry open channel bed form.

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Modeling gut neuro-epithelial connections in a novel microfluidic device

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Microsystems & Nanoengineering 2023年第6期9卷 235-246页

作者： jose M.de Hoyos-Vega Xi Yu Alan M.Gonzalez-Suarez Sisi Chen Arnaldo mercado-Perez Eugene Krueger Jeric Hernandez Yaroslav Fedyshyn Brooke R.Druliner David R.Linden Arthur Beyder Alexander Revzin Department of Physiology and Biomedical Engineering Mayo ClinicRochesterMNUSA Enteric Neuroscience Program(ENSP) Mayo ClinicRochesterMNUSA Division of Gastroenterology and Hepatology Department of MedicineMayo ClinicRochesterMNUSA

The intestinal lumen is filled with diverse chemical and physical *** epithelial cells sense these stimuli and signal to enteric neurons which coordinate a range of physiologic processes required for normal digestive tract ***,the neuro-epithelial connections remain poorly resolved,in part because the tools for orchestrating interactions between these cellular compartments are *** describe the development of a two-compartment microfluidic device for co-culturing enteric neurons with intestinal epithelial *** device contains epithelial and neuronal compartments connected by *** epithelial compartment was designed for cell seeding via injection and confinement of intestinal epithelial cells derived from human intestinal *** demonstrated that organoids planarized effectively and retained epithelial phenotype for over a *** the second chamber we dissociated and cultured intestinal myenteric neurons including intrinsic primary afferent neurons(IPANs)from transgenic mice that expressed the fluorescent protein *** extended projections into microgrooves,surrounded and frequently made contacts with epithelial *** density and directionality of neuronal projections were enhanced by the presence of epithelial cells in the adjacent *** microfluidic device represents a platform that may,in the future,be used to dissect structure and function of neuro-epithelial connections in the gut and other organs(skin,lung,bladder,and others)in health and disease.

关键词： fluid bladder organs

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Water Profile over an Ogee-Shaped Crest and the Reynolds Fractional Equations

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Journal of Mathematics and System Science 2013年第3期3卷 139-145页

作者： jose Roberto mercado Pedro Guido Aldana Waldo Ojeda Bustamante Jorge Sanchez-Sesma Mexican Institute of Water Technology Progreso Jiutepec Morelos CP 62550 Paseo Cuauhn6huac 8532 Mexico

Weir crest must have the correct shape in the concave side of an ogee-shaped crest to diminish erosion. This shape can be obtained using an approximation of the fractional Reynolds equations when the water interacts with the surface. A model is introduced for the Reynolds stresses complemented with a closure relation of fractional origin. A power type solution is obtained for the main velocity and stress. Velocity profile is found based on the assumption of a steady flow and the energy conservation equation. A Froude number and the cubic equation of the weir are built. The dimensionless upstream velocity head is also determined which allow graphically showing the exponent and coefficient of the water-profile over an ogee-shaped crest. It is possible to estimate the occupied-space index trough an exponents＇ ratio of profile over the velocity head.

关键词： Crest cornice-shaped Reynolds fractional equations water profile over crest.

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The use of high resolution anterior segment optical coherence tomography for the characterization of conjunctival lymphoma, conjunctival amyloidosis and benign reactive lymphoid hyperplasia

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Eye and Vision 2019年第1期6卷 123-130页

作者： Nandini Venkateswaran Carolina.mercado Ann Q.Tran Armando Garcia Pedro Francisco Monsalve Diaz Sander R.Dubovy Anat Galor Carol L.Karp Bascom Palmer Eye Institute University of Miami Miller School of Medicine900 NW 17th StreetMiamiFL 33136USA Department of Ophthalmology Miami Veterans Administration Medical CenterMiamiFLUSA

Background:Conjunctival lymphoma,conjunctival amyloidosis and benign reactive lymphoid hyperplasia(BRLH)are conditions that often have a similar appearance on the ocular *** use of high resolution anterior segment optical coherence tomography(HR-OCT)enables clinicians to evaluate distinctive differences in tissue morphology and cellular patterns in various ocular surface *** this study,we characterize the morphological differences seen in conjunctival lymphoma,conjunctival amyloidosis and BRLH on HR-OCT ***:A retrospective chart review was performed of patients with biopsy proven conjunctival lymphoma,conjunctival amyloidosis and BRLH between 2012 and 2019 at the Bascom Palmer Eye *** were excluded if HR-OCT imaging was not performed on initial ***:Thirty-four total eyes of 27 patients were *** eyes had conjunctival lymphoma(16 patients),8 eyes had conjunctival amyloidosis(6 patients)and 6 eyes had BRLH(5 patients).All conditions appeared clinically as pink,red or yellow subepithelial lesions but had different features on *** lymphoma,HR-OCT images typically showed homogenous,dark subepithelial lesions with smooth borders,containing monomorphic dot-like ***-OCT images of amyloidosis typically showed heterogeneous,dark lesions with irregular borders,often containing hyperreflective linear ***-OCT images of BRLH showed variable infiltration of the subepithelial tissue,at times with homogenous lesions containing dot-like infiltrates like lymphoma and other times with more hyperreflective,subepithelial *** cytometry and gene rearrangement was needed for final differentiation between BRLH and lymphoma ***:Distinctive features on HR-OCT of conjunctival lymphoma,conjunctival amyloidosis and BRLH can help characterize these lesions beyond what is apparent with the clinical *** studies can further validate this technology’s use with m

关键词： High resolution anterior segment optical coherence tomography Ocular surface imaging Ocular surface lesions Conjunctival lymphoma Conjunctival amyloidosis Benign reactive lymphoid hyperplasia

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Early Feeding vs 5-Day Fasting after Distal Elective Bowel Anastomoses in Children.A Randomized Controlled Trial

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Surgical Science 2013年第1期4卷 45-48页

作者： Roberto Davila-Perez Eduardo Bracho-Blanchet Francisco Galindo-Rocha jose Tovilla.mercado Gustavo Varela-Fascinetto Emilio Fernandez-Portilla Pablo Lezama-del-Valle Jaime Nieto-Zermeno Pediatric Surgery Department Hospital Infantil de Mexico Federico GomezMexico CityMexico

Purpose: Determine the safety and efficacy of early enteral feeding after distal elective bowel anastomoses (DEBA) in children. Methods: Controlled randomized trial including pediatric patients with DEBA, excluding non-elective and high risk patients. Variables: Demographics, operative time, anastomosis placement, beginning peristalsis and bowel movement, time to full diet intake, post-operative stay, persisting vomiting and abdominal distention, wound infection or dehiscence, anastomotic leak, reoperation, death. Randomization into: 1) Experimental group (EG): early feeding group, after a minimum 24 hours fasting period, oral fluids and diet was started;2) Control group (CG): obligatory 5-day fasting. Descriptive Statistics: Student’s t test for quantitative and Chi square for qualitative variables, a p-value 0.05 was considered significant. Results: 60 patients were included since June 2003 to May 2004, 30 ineach group. Mean age 2 years, weight16 kg, malnutrition 33%. Stomal Ethiology: Anorectal-malformation 46%, Hirschsprung 13%, inflammatory 35%, tumoral 5%. Mostly in colon 71%. Mean surgical time 142 min. None developed vomiting or required nasogastric-tube. Mild abdominal distension 13%, mild fever 16.5% and wound complications 18%. Anastomosis leakage 6.5%, none required reoperations. Demographic variables showed no statistical differences. Full oral intake was in the 2nd postoperative day in the EG vs CG (p = 0.001). Postoperative hospital stay was 6.0 ± 3 in the EG vs 9.8 ± 4 days in the CG with clinical but not statistical significance. Peristalsis beginning, first flatus passage and bowel movements showed no statistical differences. The complication incidence was low and equally distributed. Conclusions: Early feeding after DEBA is safe and well tolerated in children.

关键词： Early Feeding Bowel Anastomosis Children

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The Bovine Tuberculosis Campaign in Mexico： Present and Perspectives

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Journal of Agricultural Science and Technology(A) 2014年第4期4卷 277-290页

作者： jose Alfredo Gutierrez-Reyes Estela Flores-Velazquez Guillermo Agustin Reyes-Escalona Joaquin BraulioDelgadillo-Alvarez Marcela Amalia.mercado-Pezzat Eli Tonatiuh Selva-Hernandez Alejandro Perera-Ortiz jose Alberto Ordufia-Sumaran Cecilia Romero-Torres Luis Antonio Jimenez-Zamudio Ethel AwildaGarcfa-Latorre2 and Citlatepetl Salinas-Lara General Directorate of Animal Health National Service of Health Food Safety and Food Quality (SENASICA) Mexico City 03310 Mexico National School of Biological Sciences National Polythecnical Institute Santo Tomas Unit Lazaro Cardenas Mexico City 11340 Mexico

Mexico is considered by different international organizations as a country where the bovine tuberculosis is an endemic problem. However, there are important and consistent improvements in eradicating the disease by the implementation of government policies and with the livestock organization support. The current situation of the program shows good results regarding decreasing the prevalence, and as a consequence, the risk of this disease for public health and animal health, as well as the improvement in animal production and international and national trade. Today, the bovine tuberculosis distribution is almost known in Mexico. In addition, a series of controls and indicators have been established, which allow maintain consistency and reliability in the data that are managed and analyzed.

关键词： Bovine tuberculosis surveillance status quarantine Mexico.

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Cross-Linked Alginate Film Pore Size Determination Using Atomic Force Microscopy and Validation Using Diffusivity Determinations

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Journal of Surface Engineered Materials and Advanced Technology 2013年第4期3卷 1-12页

作者： Cheryl Simpliciano Larissa Clark Behrokh Asi Nathan Chu Maria.mercado Steven Diaz Michel Goedert Maryam Mobed-Miremadi Department of Biomedical Chemical and Materials Engineering San Jose State University San Jose USA.

The deficit of organ donors has fueled the need for advances in tissue engineering and regenerative medicine. Microencapsulation in alginate immuno-isolation membranes has been used to treat many disabling metabolic disorders, namely, phenylketonuria, kidney failure and diabetes mellitus. Systematic nutrient flux determinations are hindered by the lack of experimental data on alginate-based membrane topography and the pore size thus preventing the full therapeutic potential of the bio-membranes to be reached. In this study, samples of cross-linked alginate membranes were subjected to the following analytical characterization: 1) pore size characterization using atomic force microscopy operated in contact mode to detect and measure pore size;2) differential scanning calorimetry to confirm biopolymer cross-linking;and 3) diffusivity measurements using spectrophotometry and fluorescence microscopy to confirm the presence of through pores and to calculate reflection coefficients. The pore sizes for the pre-clinical standard formulation of 1.5% (w/v) medium viscosity alginate cross-linked with 1.5% CaCl2 and 0.5% (w/v) alginate and chitosan cross-linked with 20% CaCl2 are 5.2 nm ± 0.9 nm and 7.0 nm ± 3.1 nm, respectively. An increase in the glass transition temperatures as a function of cross-linker concentration was observed. Diffusivity values obtained from the inward diffusivity of creatinine into macrocapsules (d = 1000 μm ± 75 μm) and the outward diffusivity of FITC dextrans from macrocapsules (d = 1000 μm ± 75 μm) and microcapsules (d = 40 μm ± 5 μm) were shown to correlate strongly (R2 = 0.9835) with the ratio of solute to pore sizes, confirming the presence of through pores. Reflection coefficients approaching and exceeding unity correlate with the lack of permeability of the membranes to MW markers that are 70 kDa and greater.

关键词： Alginate Atomic Force Microscopy Pore Size Stokes’ Radius Diffusivity Cross-linking Differential Scanning Calorimetry Reflection Coefficient

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Turbo-taxonomy to assemble a megadiverse lichen genus:seventy new species of Cora(Basidiomycota:Agaricales:Hygrophoraceae),honouring David Leslie Hawksworth’s seventieth birthday

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Fungal Diversity 2017年第3期84卷 139-207页

作者： Robert Lücking Manuela Dal Forno Bibiana Moncada Luis Fernando Coca Leidy Yasmín Vargas-Mendoza André Aptroot Laura Juliana Arias Betty Besal Frank Bungartz Diego Mauricio Cabrera-Amaya Marcela E.S.Cáceres José Luis Chaves Sionara Eliasaro Martha Cecilia Gutiérrez Jesús E.Hernández Marin María de los Ángeles Herrera-Campos María E.Holgado-Rojas Harald Jonitz Martin Kukwa Fabiane Lucheta Santiago Madriñán Marcelo Pinto Marcelli Suzana Maria de Azevedo Martins Joel a.mercado-Díaz Jorge Alberto Molina Eduardo A.Morales Peter R.Nelson Freddy Nugra Francisco Ortega Telma Paredes Ayda Lucía Patiño Rouchi Nadine Peláez-Pulido Rosa Emilia Pérez Pérez Gary B.Perlmutter Eimy Rivas-Plata Javier Robayo Camilo Rodríguez Diego Fernando Simijaca Edier Soto-Medina Adriano Afonso Spielmann Alejandra Suárez-Corredor Jean-Marc Torres Carlos Alberto Vargas Alba Yánez-Ayabaca Gothamie Weerakoon Karina Wilk Marcela Celis Pacheco Mauricio Diazgranados Grischa Brokamp Thomas Borsch Patrick M.Gillevet Masoumeh Sikaroodi James D.Lawrey Botanical Garden and Botanical Museum Berlin KöniginLuise-Straße 6-814195 BerlinGermany Science Education The Field Museum1400 South Lake ShoreChicagoIL 60605USA Department of Botany Smithsonian InstitutionNational Museum of Natural History10th St Constitution Ave NWWashingtonDC 20560USA Department of Environmental Science and Policy George Mason UniversityFairfaxVA 22030-4444USA Licenciatura en Biologı´a Universidad Distrital Francisco Jose´de CaldasCra.4 No.26D-54Torre de LaboratoriosHerbarioBogota´D.C.Colombia Herbario Universidad de Caldas(FAUC) Edificio BicentenarioCalle 65No.26-10ManizalesA.A.275CaldasColombia ABL Herbarium G.v.d.Veenstraat 1073762 XK SoestThe Netherlands Biologı´a Universidad del Quindı´oCarrera 15 Calle 12 NorteArmeniaQuindı´oColombia 120 Chavis Avenue LexingtonVA 24450USA Charles Darwin Foundation Puerto AyoraSanta Cruz IslandGala´pagosEcuador Schedestraße 8 53113 BonnGermany Jardı´n Bota´nico de Bogota´jose´Celestino Mutis Avenida Calle 63 No.68-95Bogota´D.C.Colombia Departamento de Biocieˆncias Universidade Federal de SergipeAvenida Vereador Olı´mpio Grandes/n.ItabaianaCEP:49.500-000 SergipeBrazil Laboratorio de Hongos Instituto Nacional de Biodiversidad(INBio)Apdo.22-3100Santo Domingo de HerediaCosta Rica Departamento de Botaˆnica Universidade Federal do Parana´Campus Polite´cnicoSetor de Cieˆncias Biolo´gicasAv.Cel.Francisco H.dos Santoss/nJardim das Ame´ricas81531-980 CuritibaPRBrazil Ingenierı´a Ambiental Universidad Distrital Francisco Jose´de CaldasCarrera 5 Este No.15-82Av.CircumvalarBogota´D.C.Colombia Instituto Experimental Jardı´n Bota´nico Dr.Tobı´as Lasser Centro de Investigacio´n y DesarrolloUniversidad Central de VenezuelaAve.Salvador AllendeJardı´n Bota´nico de CaracasApartado#2156Caracas 1010-AVenezuela Departamento de Bota´nica Instituto de Biologı´aUniversidad Nacional Auto´noma de Me´xicoApartado Postal 70-36704510 Ciudad De Me´xicoMexico Facultad de Ciencias Escuela Profesional de Biologı´aUniv

Following a large-scale phylogenetic study of the lichenized genus Cora(Basidiomycota:Agaricales:Hygrophoraceae),we formally describe 70 new species,honouring the seventieth birthday of David Leslie Hawksworth,one of the preeminent figures in mycology and lichenology in the past 50 *** on an updated phylogeny using the ITS fungal barcoding locus,we now recognize 189 taxa in a genus that until recently was considered to represent a single species;including this contribution,92 of these are formally recognized,including five taxa based on historical names or collections that have not been *** of Cora can be recognized by a combination of morphological(size,colour,lobe configuration,surface hairs,hymenophore size and shape),anatomical(thallus thickness,cortex structure,photobiont type,hyphal papillae),and ecogeographical features(substrate,habitat,distribution),and a keytable allowing the identification of all accepted taxa is *** new species are:Cora accipiter Moncada,Madrin˜a´n&Lücking ***.,*** Moncada,Soto-Medina&Lücking ***.,*** Moncada,Dal Forno&Lücking ***.,*** Dal Forno,Chaves&Lücking ***.,*** Moncada,***ı´guez&Lücking ***.,*** Lücking,Moncada&*** ***.,*** Moncada,Ya´nez-Ayabaca&Lücking ***.,*** Moncada,Patin˜o&Lücking ***.,*** Lücking,***&Dal Forno ***.,*** Holgado,Rivas Plata&Perlmutter ***.,*** Dal Forno,Eliasaro&Spielmann ***.,*** Nugra,Dal Forno&Lücking ***.,*** Lücking,Martins&Lucheta ***.,*** Moncada,R.-***´rez&Lücking ***.,*** Moncada,***.&Lücking ***.,*** Moncada,CabreraAmaya&Lücking ***.,*** Moncada,R.-***´rez&***.,*** Lücking,***&Dal Forno ***.,*** Moncada,***´rez-Corredor&Lücking ***.,*** Moncada,***&Lücking ***.,***

关键词： Bolivia Citizen science Chile Mexico Puerto Rico Sri Lanka

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