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The spectrum of building block conformers sustains the biophysical properties of clinically-oriented self-assembling protein nanoparticles

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Science China materials 2022年第6期65卷 1662-1670页

作者： Eric Voltà-Durán Julieta m Sánchez Hèctor-López-Laguna Eloi Parladé laura Sánchez-García Alejandro Sánchez-Chardi Ario de marco Ugutz Unzueta Esther Vázquez Antonio Villaverde Institut de Biotecnologia i de Biomedicina Universitat Autònoma de BarcelonaBellaterra 08193BarcelonaSpain Departament de Genètica i de microbiologia Universitat Autònoma de BarcelonaBellaterra 08193BarcelonaSpain CIBER de Bioingeniería Biomateriales y Nanomedicina(CIBER-BBN) Bellaterra 08193BarcelonaSpain Instituto de Investigaciones Biológicas y Tecnológicas(IIBYT)(CONICET-Universidad Nacional de Córdoba) ICTA&Cátedra de Química BiológicaDepartamento de QuímicaFCEFyNUNC.Av.Velez Sarsfield 1611X 5016GCA CórdobaArgentina Servei de microscòpia Universitat Autònoma de BarcelonaBellaterra 08193BarcelonaSpain Departament de Biologia Evolutiva Ecologia i Ciències AmbientalsFacultat de BiologiaUniversitat de Barcelona08028 BarcelonaSpain Laboratory for Environmental and Life Sciences University of Nova GoricaNova Gorica 5000Slovenia Biomedical Research Institute Sant Pau(IIB Sant Pau) Sant Antoni Ma Claret 16708025 BarcelonaSpain

Histidine-rich peptides confer self-assembling properties to recombinant proteins through the supramolecular coordination with divalent *** fact allows the cost-effective,large-scale generation of microscopic and macroscopic protein materials with intriguing biomedical *** such materials,resulting from the simple bioproduction of protein building blocks,homomeric nanoparticles are of special value as multivalent interactors and drug ***,we have here identified that the assembly of a given His-tagged protein might render distinguishable categories of self-assembling protein *** fact has been scrutinized through the nanobody-containing fusion proteins Em1-GFP-H6 and A3C8-GFP-H6,whose biosynthesis results in two distinguishable populations of building *** one of them,the assembling and disassembling is controllable by ***,a second population immediately self-assembles upon purification through a non-regulatable pathway,rendering larger nanoparticles with specific biological *** structural analyses of both model proteins and nanoparticles revealed important conformational variability in the building *** fact renders different structural and functional categories of the final soft materials resulting from the participation of energetically unstable intermediates in the oligomerization *** data illustrate the complexity of the Hismediated protein assembling in recombinant proteins but they also offer clues for a better design and refinement of protein-based nanomedicines,which,resulting from biological fabrication,show an architectonic flexibility unusual among biomaterials.

关键词： recombinant proteins self-assembling protein materials nanoparticles biomaterials

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Arachidyl amido cholanoic acid improves liver glucose and lipid homeostasis in nonalcoholic steatohepatitis via AmPK and mTOR regulation

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World Journal of Gastroenterology 2020年第34期26卷 5101-5117页

作者： David Fernández-Ramos Fernando Lopitz-Otsoa laura Delacruz-Villar Jon Bilbao martina Pagano laura mosca maider Bizkarguenaga marina Serrano-macia mikel Azkargorta marta Iruarrizaga-Lejarreta Jesús Sot Darya Tsvirkun Sebastiaan martijn van Liempd Felix m Goni Cristina Alonso maría Luz martínez-Chantar Felix Elortza Liat Hayardeny Shelly C Lu Josém mato Precision medicine and metabolism Laboratory Centro de Investigación Cooperativa en Biociencias(CIC bioGUNE)Derio 48160BizkaiaSpain CIBERehd-Centro de Investigación Biomédica en Red de enfermedades hepáticas y digestivas Madrid 28029Spain Department of Precision medicine University of Campania"Luigi Vanvitelli"Naples 80138Italy Liver Disease Laboratory Centro de Investigación Cooperativa en Biociencias(CIC bioGUNE)Derio 48160Spain Proteomics Platform Centro de Investigación Cooperativa en Biociencias(CIC bioGUNE)Derio 48160Spain OWL metabolomics Derio 48160Spain Instituto Biofisika(UPV/EHU CSIC)Leioa 48940SpainDepartamento de Bioquímica y Biología MolecularUniversidad del País VascoLeioa 48940Spain Pre-clinical and Chemistry Manufacturing and ControlsGalmed PharmaceuticalsTel Aviv 6578317Israel metabolomics Platform Centro de Investigación Cooperativa en Biociencias(CIC bioGUNE)Derio 48160Spain Division of Digestive and Liver Diseases Cedars-Sinai Medical CenterLos AngelesCA 90048United States

BACKGROUND Arachidyl amido cholanoic acid(Aramchol)is a potent downregulator of hepatic stearoyl-CoA desaturase 1(SCD1)protein expression that reduces liver triglycerides and fibrosis in animal models of *** a phase IIb clinical trial in patients with nonalcoholic steatohepatitis(NASH),52 wk of treatment with Aramchol reduced blood levels of glycated hemoglobin A1c,an indicator of glycemic *** To assess lipid and glucose metabolism in mouse hepatocytes and in a NASH mouse model[induced with a 0.1%methionine and choline deficient diet(0.1mCD)]after treatment with *** Isolated primary mouse hepatocytes were incubated with 20μmol/L Aramchol or vehicle for 48 ***,analyses were performed including Western blot,proteomics by mass spectrometry,and fluxomic analysis with 13C-uniformly labeled *** the in vivo part of the study,male C57BL/6J mice were randomly fed a control or 0.1mCD for 4 wk and received 1 or 5 mg/kg/d Aramchol or vehicle by intragastric gavage for the last 2 *** metabolomics were assessed using ultra-high-performance liquid chromatography-time of flight-mS for the determination of glucose metabolism-related *** Combination of proteomics and Western blot analyses showed increased AmPK activity while the activity of nutrient sensor mTORC1 was decreased by Aramchol in *** translated into changes in the content of their downstream targets including proteins involved in fatty acid(FA)synthesis and oxidation[PACCα/β(S79),SCD1,CPT1A/B,HADHA,and HADHB],oxidative phosphorylation(NDUFA9,NDUFB11,NDUFS1,NDUFV1,ETFDH,and UQCRC2),tricarboxylic acid(TCA)cycle(mDH2,SUCLA2,and SUCLG2),and ribosome(P-p70S6K[T389]and P-S6[S235/S236]).Flux experiments with 13Cuniformely labeled glucose showed that TCA cycle cataplerosis was reduced by Aramchol in hepatocytes,as indicated by the increase in the number of rounds that malate remained in the TCA ***,liver metabolomic analysis showed

关键词： Nonalcoholic fatty liver disease Steatohepatitis methionine and choline deficient diet Tricarboxylic acid cycle Hemoglobin A1c Stearoyl-CoA desaturase 1

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ABCB1 and ABCB19 auxin transporters have synergistic effects on early and late Arabidopsis anther development

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Journal of Integrative Plant Biology 2015年第12期57卷 1089-1098页

作者： Valentina Cecchetti Patrizia Brunetti Nadia Napoli laura Fattorini maria maddalena Altamura Paolo Costantino maura Cardarelli IBPm-CNR Institute of molecular Biology and Pathology National Research Council Sapienza University of Rome Department of Biology and Biotechnology Sapienza University of Rome Department of Environmental Biology Sapienza University of Rome

Arabidopsis abcbl abcb19 double mutants defective in the auxin transporters ABCB1/PGP1 and ABCB19/PGP19 are altered in stamen elongation, anther dehiscence and pollen maturation. To assess the contribution of these transporters to stamen development we performed phenotypic, histological analyses, and in situ hybridizations on abcbl and abcbl9 single mutant flowers. We found that pollen maturation and anther dehiscence are precocious in the abcbl but not in the abcb19 mutant. Accordingly, endothecium ligniflcation is altered only in abcbl anthers. Both abcbl and abcb1 abcb19 stamens also show altered early development, with asynchronous anther Iocules and a multilayer tapetum. DAPI staining showed that the timing of meiosis is asynchronous in abcbl abcb19 anther Iocules, while only a small percentage of pollen grains are non- viable according to Alexander＇s staining. In agreement, TAm （TARDY ASYNCHRONOUS mEIOSIS）, as well as BAm2 （BARELY ANY mERISTEm）involved in tapetal cell development--areoverexpressed in abcbl abcb19 young flower buds. Corre- spondingly, ABCB1 and ABCB19 mRNA localization supports the observed phenotypes of abcbl and abcbl abcb19 mutant anthers. In conclusion, we provide evidence that auxin transport plays a significant role both in early and late stamen development： ABCB1 plays a major role during anther development, while ABCB19 has a synergistic role.

关键词： Anther development Arabidopsis auxin transport endothecium lignification tapetum differentiation

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metabolic shift in liver: Correlation between perfusion temperature and hypoxia inducible factor-1α

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World Journal of Gastroenterology 2015年第4期21卷 1108-1116页

作者： Andrea Ferrigno laura Giuseppina Di Pasqua Alberto Bianchi Plinio Richelmi mariapia VairettiAndrea Ferrigno Unit of Cellular and molecular Pharmacology and Toxicology Department of Interna Medicine and TherapeuticsUniversity of PaviaPavia 27100 Italy

AIm: To study at what temperature the oxygen carried by the perfusate meets liver requirements in a model of organ perfusion. mETHODS: in this study, we correlated hypoxia induciblefactor(Hi F)-1α expression to the perfusion temperature and the hepatic oxygen uptake in a model of isolated perfused rat liver. Livers from Wistar rats were perfused for 6 h with an oxygenated medium at 10, 20, 30 and 37 ℃. Oxygen uptake was measured by an oxygen probe; lactate dehydrogenase activity, lactate release and glycogen were measured spectrophotometrically; bile flow was gravitationally determined; p H of the perfusate was also evaluated; Hi F-1α m RNA and protein expression were analyzed by real time-polymerase chain reaction and ELi SA, respectively. RESULTS: Livers perfused at 10 and 20 ℃ showed no difference in lactate dehydrogenase release after 6 h of perfusion(0.96 ± 0.23 vs 0.93 ± 0.09 m U/min per g) and had lower hepatic damage as compared to 30 and 37 ℃(5.63 ± 0.76 vs 527.69 ± 45.27 m U/min per g, respectively, P s mol/mg, respectively, P s mol/m L, respectively, P s m RNA(0.40 ± 0.08 and 0.20 ± 0.03 vs 0.60 ± 0.20 and 1.47 ± 0.30, respectively, P s < 0.05) and protein(3.72 ± 0.16 and 3.65 ± 0.06 vs 4.43 ± 0.41 and 6.44 ± 0.82, respectively, P s < 0.05) ***: Livers perfused at 10 and 20 ℃ show no sign of liver injury or anaerobiosis, in contrast to livers perfused at 30 and 37 ℃.

关键词： Anaerobiosis Hypoxia inducible factor-1 α ischemia

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miniscope3D:optimized single-shot miniature 3D fluorescence microscopy

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Light(Science & Applications) 2020年第1期9卷 367-379页

作者： Kyrollos Yanny Nick Antipa William Liberti Sam Dehaeck Kristina monakhova Fanglin Linda Liu Konlin Shen Ren Ng laura Waller UCB/UCSF Joint Graduate Program in Bioengineering University of CaliforniaBerkeleyCA 94720USA Department of Electrical Engineering&Computer Sciences University of CaliforniaBerkeleyCA 94720USA TIPs Department Universite libre de Bruxelles(ULB)1050 BrusselsBelgium

miniature fluorescence microscopes are a standard tool in systems ***,widefield miniature microscopes capture only 2D information,and modifications that enable 3D capabilities increase the size and weight and have poor resolution outside a narrow depth ***,we achieve the 3D capability by replacing the tube lens of a conventional 2D miniscope with an optimized multifocal phase mask at the objective’s aperture *** the phase mask at the aperture stop significantly reduces the size of the device,and varying the focal lengths enables a uniform resolution across a wide depth *** phase mask encodes the 3D fluorescence intensity into a single 2D measurement,and the 3D volume is recovered by solving a sparsity-constrained inverse *** provide methods for designing and fabricating the phase mask and an efficient forward model that accounts for the fieldvarying aberrations in miniature *** demonstrate a prototype that is 17mm tall and weighs 2.5 grams,achieving 2.76μm lateral,and 15μm axial resolution across most of the 900×700×390μm^(3) volume at 40 volumes per *** performance is validated experimentally on resolution targets,dynamic biological samples,and mouse brain *** with existing miniature single-shot volume-capture implementations,our system is smaller and lighter and achieves a more than 2×better lateral and axial resolution throughout a 10×larger usable depth *** microscope design provides single-shot 3D imaging for applications where a compact platform matters,such as volumetric neural imaging in freely moving animals and 3D motion studies of dynamic samples in incubators and lab-on-a-chip devices.

关键词： enable resolution narrow

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Three-dimensional fuzzy graphene ultra-microelectrodes for subcellular electrical recordings

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Nano Research 2020年第5期13卷 1444-1452页

作者： Sahil K.Rastogi Jacqueline Bliley laura matino Raghav Garg Francesca Santoro Adam W.Feinberg Tzahi Cohen-Karni Department of Biomedical Engineering Carnegie Mellon UniversityPittsburghPA15213USA Center for Advanced Biomaterials for Healthcare Istituto Italiano di TecnologiaNaples80125Italy Department of Chemical Materials and Industrial Production EngineeringUniversity of Naples Federico IINaples80125Italy Department of materials Science and Engineering Carnegie Mellon UniversityPittsburghPA15213USA

microelectrode arrays(mEAs)have enabled investigation of cellular networks at sub-millisecond temporal ***,current mEAs are limited by the large electrode footprint since reducing the electrode’s geometric area to sub-cellular dimensions leads to a significant increase in impedance thus affecting its recording *** report a breakthrough ultra-microelectrodes platform by leveraging the outstanding surface-to-volume ratio of nanowire-templated out-of-plane synthesized three-dimensional fuzzy graphene(NT-3DFG).The enormous surface area of NT-3DFG leads to 140-fold reduction in electrode impedance compared to bare Au microelectrodes,thus enabling scaling down the geometric size by 625-fold to ca.2µm×2µ*** out-of-plane morphology of NT-3DFG leads to a tight seal with the cell membrane thus enabling recording of electrical signals with high signal-to-noise ratio(SNR)of>*** work highlights the possibility to push the limits of the conventional mEA technology to enable electrophysiological investigation at sub-cellular level without the need of any surface *** presented approach would greatly impact our basic understanding of signal transduction within a single cell as well as complex cellular assemblies.

关键词： microelectrode array ultra-microelectrodes electrical recordings biocompatible hybrid-nanomaterial 3D graphene cardiomyocytes

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An augmented and interactive exhibition of an archived model for Frederick Kiesler's Endless House, 1959

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Frontiers of Architectural Research 2022年第6期11卷 993-1006页

作者： Chantelle Niblock laura mcGuire John Harding Gerd Zillner Chris Hamill Andrew Whitney School of Natural and Built Environment Queens University of BelfastNorthern IrelandBT71NNUK School of Architecture University of Hawaii at MānoaHonoluluHI96822USA School of Architecture University of ReadingReadingBerkshireRG66URUK Austrian Frederick and Lillian Kiesler Private Foundation Mariahilfer Straße1060WienAustria Hotknife Digital media NottinghamNG13HDUK

This research explores the potential of an immersive and interactive online archive to enhance our understanding of historic architecture through the study of *** reports on implementing an augmented reality mobile application that exhibits a model for the unbuilt Endless House,1959,by Frederick Kiesler.A reflective critique,from the researcher’s point of view,and initial feedback from a small sample of architecture students,provides an insight into users’experience of the exhibition,its value as a research tool,and as an educational *** on existing technologies and established research methods,we present an alternative way of exhibiting a large-scaled model for public engagement and research collaboration between academics,archivists,and *** discuss the development of the mobile application with interactive features specifically designed for an architectural *** touches on issues associated with documenting,interpreting,and exhibiting architectural models,emphasizing accessibility,accuracy,engagement,combining 3D and 2D digital assets,and user *** was found that the interactive and immersive features of the exhibition enhanced the researchers’scope to spatially inspect the model,visually experience it,collaborate with others,and strengthen connections between the model and other examples of Kiesler’s textual and visual archival materials.

关键词： Augmented model Unbuilt architecture Frederick Kiesler Interactive exhibition Endless house

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Neurofilament light chain is increased in the parahippocampal cortex and associates with pathological hallmarks in Parkinson’s disease dementia

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Translational Neurodegeneration 2023年第1期12卷 872-888页

作者： Irene Frigerio max A.Laansma Chen-Pei Lin Emma J.m.Hermans maud m.A.Bouwman John G.J.m.Bol Yvon Galis-de Graaf Dagmar H.Hepp Annemieke J.m.Rozemuller Frederik Barkhof Wilma D.J.van de Berg laura E.Jonkman Section Clinical Neuroanatomy and Biobanking Department of Anatomy and NeurosciencesAmsterdam UMC Location Vrije Universiteit AmsterdamDe Boelelaan 1118AmsterdamThe Netherlands Amsterdam Neuroscience NeurodegenerationAmsterdamThe Netherlands Amsterdam Neurosci-ence Brain ImagingAmsterdamThe Netherlands Department of Neurology Amsterdam UMC Location Vrije Universiteit AmsterdamDe Boelelaan 1117AmsterdamThe Netherlands Department of Pathology Amsterdam UMC Location Vrije Universiteit AmsterdamDe Boelelaan 1117AmsterdamThe Netherlands Department of Radiology and Nuclear medicine Amsterdam UMC Location Vrije Universiteit AmsterdamDe Boelelaan 1117AmsterdamThe Netherlands Institutes of Neurology and Healthcare Engineering University College LondonLondonUK.

Background Increased neurofilament levels in biofluids are commonly used as a proxy for neurodegeneration in several neurodegenerative *** this study,we aimed to investigate the distribution of neurofilaments in the cerebral cortex of Parkinson’s disease(PD),PD with dementia(PDD)and dementia with Lewy bodies(DLB)donors,and its association with pathology load and mRI measures of atrophy and *** Using a within-subject post-mortem mRI-pathology approach,we included 9 PD,12 PDD/DLB and 18 age-matched control *** thickness and mean diffusivity(mD)metrics were extracted respectively from 3DT1 and DTI at 3T in-situ *** autopsy,pathological hallmarks(pSer129-αSyn,p-tau and amyloid-βload)together with neurofilament light-chain(NfL)and phosphorylated-neurofilament medium-and heavy-chain(p-Nfm/H)immunoreactivity were quantified in seven cortical regions,and studied in detail with confocal-laser scanning *** correlations between mRI and pathological measures were studied using linear mixed *** Compared to controls,p-Nfm/H immunoreactivity was increased in all cortical regions in PD and PDD/DLB,whereas NfL immunoreactivity was increased in the parahippocampal and entorhinal cortex in PDD/***-positive neurons showed degenerative morphological features and axonal *** increased p-Nfm/H correlated with p-tau load,and NfL correlated with pSer129-αSyn but more strongly with p-tau load in PDD/***,neuro-filament immunoreactivity correlated with cortical thinning in PD and with increased cortical mD in PDD/*** Taken together,increased neurofilament immunoreactivity suggests underlying axonal injury and neurofilament accumulation in morphologically altered neurons with increased pathological ***,we demonstrate that such neurofilament markers at least partly explain mRI measures that are associated with the neurodegenerative process.

关键词： Neurofilament NfL Axonal degeneration Parkinson’s disease Parkinson’s disease dementia Dementia with Lewy bodies Cortical thickness Cortical atrophy mean diffusivity

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Cellular crosstalk during cholestatic liver injury

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Liver Research 2017年第1期1卷 26-33页

作者： Joanne Thomson laura Hargrove Lindsey Kennedy Jennifer Demieville Heather Francis Research Central Texas Veterans Healthcare SystemTXUSA Digestive Disease Research Center Baylor Scott&White HealthTXUSA medicine Texas A&M Health Science CenterTempleTXUSA

The functions of the liver are very *** detoxifying blood to storing glucose in the form of glycogen and producing bile to facilitate fat digestion,the liver is a very active and important *** liver is comprised of many varied cell types whose functions are equally *** line the biliary tree and aid in transporting and adjusting the composition of bile as it travels to the *** stellate cells and portal fibroblasts are located in different areas within the liver architecture,but both contribute to the development of fibrosis upon activation after liver *** cells,including those that constitute the peribiliary vascular plexus,are involved in functions other than blood delivery to and from the liver,such as supporting the growth of the biliary tree during *** cells are normally found in healthy livers but in very low ***,after injury,mast cell numbers greatly increase as they infiltrate and release factors that exacerbate the fibrotic *** not an all-inclusive list,these cells have individual roles within the liver,but they are also able to communicate with each other by cellular *** this review,we examine some of these pathways that can lead to an increase in the homeostatic dysfunction seen in liver injury.

关键词： mast cells Paracrine Cholangiocytes Hepatic stellate cells Crosstalk

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Effects of combined pre- and post-natal protein deprivation on the myenteric plexus of the esophagus of weanling rats:A histochemical,quantitative and ultrastructural study

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World Journal of Gastroenterology 2007年第26期13卷 3598-3604页

作者： Edson A Liberti Ricardo BV Fontes Verginia m Fuggi laura Bm maifrino Romeu R Souza Autonomic Nervous System Laboratory (VQm) Department of AnatomyInstitute of Biomedical SciencesUniversity of So Paulo So PauloBrazil

AIm: To evaluate the effects of protein deprivation on the myenteric plexus of the esophagus of weanling rats. mETHODS: Pregnant female Wistar rats were divided into 2 groups: nourished (N),receiving normal diet,and undernourished (D),receiving a protein-deprived diet,which continued after birth. At twenty-one days of age,13 esophagi from each group were submitted to light microscopy and morphometrical analysis employing the NADH diaphorase,NADPH diaphorase and acetylcholinesterase techniques. Three other esophagi from each group were evaluated with transmission electron microscopy (TEm). RESULTS: In both the NADH- and the NADPH-reactive mounts,the neurons of the N mounts were more intensely stained,while in the D esophagi only the larger neurons were reactive. many myenteric neurons of N were intensely reactive for AChE activity but only a few neurons of D exhibited these aspects. Ultrastructural analysis revealed that the granular reticulum of N showed large numbers of ribosomes aligned on the outer surface of its regularly arranged membrane while the ribosomes of D were disposed in clusters. The chromatin was more homogeneously scattered inside the neuron nucleus of N as well as the granular component of the nucleolus was evidently more developed in this group. Statistically significant differences between N and D groups were detected in the total estimated number of neurons stained by the NADPH technique. CONCLUSION: The morphological and quantitative data shows that feeding with protein-deprived diet in 21-d old rats induces a delay in the development of the myenteric neurons of the esophagus.

关键词： Enteric nervous system myenteric plexus Nutrition microscopy Electron Transmission Esopha-gus Rat Wistar

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