Understanding the interaction mechanisms of engineered nanomaterials(ENMs)with plant membranes is crucial for their effective use in various applications.While passive transport of smaller ENMs is well-documented,the ...Understanding the interaction mechanisms of engineered nanomaterials(ENMs)with plant membranes is crucial for their effective use in various applications.While passive transport of smaller ENMs is well-documented,the mechanisms underlying active transport of larger ENMs remain poorly understood.This study systematically investigates the active transport and subcellular distribution of ENMs(100-1000 nm)within protoplasts using optical ratiometric silica pH sensors for localization.Highly monodispersed ratiometric pH sensors,based on silica particles modified with fluorescein-5-isothiocyanate(FITC)and cyanine3 NHS ester(CY3)dyes,were employed to elucidate internalization mechanisms.Protoplasts from Nicotiana tabacum L.leaves successfully internalized the sensors.3D segmentation of protoplasts revealed distinct pH gradients,indicating vacuole accumulation.Colocalization analysis and cellular compartments staining further confirmed sensor distribution.High-throughput imaging flow cytometry showed efficient internalization rates,which decreased after cell wall regeneration.Notably,inhibition experiments with the salicylic acid(SA)and Tyrphostin A23(TyrA23)inhibitors confirmed clathrin-mediated endocytosis in particle uptake.This study establishes rational design principles for controlling active ENM uptake and subcellular localization via optical pH sensing in protoplasts.The findings enhance our understanding of plant cell trafficking mechanisms and hold promise for targeted delivery and applications in plant biology research.展开更多
The shortage of tissues and organs for transplantation is an urgent clinical concern.In situ 3D printing is an advanced 3D printing technique aimed at printing the new tissue or organ directly in the patient.The ink f...The shortage of tissues and organs for transplantation is an urgent clinical concern.In situ 3D printing is an advanced 3D printing technique aimed at printing the new tissue or organ directly in the patient.The ink for this process is central to the outcomes,and must meet specific requirements such as rapid gelation,shape integrity,stability over time,and adhesion to surrounding healthy tissues.Among natural materials,silk fibroin exhibits fascinating properties that have made it widely studied in tissue engineering and regenerative medicine.However,further improvements in silk fibroin inks are needed to match the requirements for in situ 3D printing.In the present study,silk fibroin-based inks were developed for in situ applications by exploiting covalent crosslinking process consisting of a pre-photo-crosslinking prior to printing and in situ enzymatic crosslinking.Two different silk fibroin molecular weights were characterized and the synergistic effect of the covalent bonds with shear forces enhanced the shift in silk secondary structure towardβ-sheets,thus,rapid stabilization.These hydrogels exhibited good mechanical properties,stability over time,and resistance to enzymatic degradation over 14 days,with no significant changes over time in their secondary structure and swelling behavior.Additionally,adhesion to tissues in vitro was demonstrated.展开更多
Sorghum （Sorghum bicolor） has high levels of starch, sugar, and fiber and is one of the most important energy crops in the world. Insect damage is one of the challenges that impacts sorghum biomass production. There...Sorghum （Sorghum bicolor） has high levels of starch, sugar, and fiber and is one of the most important energy crops in the world. Insect damage is one of the challenges that impacts sorghum biomass production. There are at least 150 insect species that can infest sorghum varieties worldwide. These insects can complete several generations within a growing season, they target various parts of sorghum plants at devel- opmental stages, and they cause significant biomass losses. Genetic research has revealed the existence of resistant genetics in sorghum and insect tolerant sorghum varieties have been identified. Various control methods have been developed, yet more effective management is needed for increasing sorghum biomass production. Although there are no transgenic sorghum products on the market yet, biotechnology has been recognized as an important tool for controlling insect pests and increasing sorghum production.展开更多
In 1986, Binnig et al. developed the first atomic force microscope (AFM). The AFM, unlike the scanning tunnelling microscope (STM), has no demands for electrical conductivity, so it has been used in science and techno...In 1986, Binnig et al. developed the first atomic force microscope (AFM). The AFM, unlike the scanning tunnelling microscope (STM), has no demands for electrical conductivity, so it has been used in science and technology more widely. In 1988, the AFM was improved, and the AFM employing laser beam deflection for force detection (laser-AFM) was developed. In 1990, laser-AFM got the atomic-resolution. Up till now, the AFM has developed into a very important technique for studying the surface.展开更多
The principle of scanning probe microscopes (SPM) was lust described by J. A. O’Keefe in the 1960s. In 1982, the scanning tunnelling microscope (STM), the first supreme example of SPM family, was developed; for which...The principle of scanning probe microscopes (SPM) was lust described by J. A. O’Keefe in the 1960s. In 1982, the scanning tunnelling microscope (STM), the first supreme example of SPM family, was developed; for which Binnig and Rohrer received the 1986 Nobel Prize in Physics. Shortly after that, in 1986 Binnig together with Quate and Gerber introduced the first atomic force microscope (AFM). Unlike the STM, the AFM展开更多
基金supported by the European Research Council(ERC)under the European Union’s Horizon 2020 research and innovation program ERC Starting Grant“INTERCELLMED”(Nr.759959)the Associazione Italiana per la Ricerca contro il Cancro(AIRC)(MFAG-2019,Nr.22902)+4 种基金the PRIN 2022(Nr.2022CRFNCP_PE11_PRIN2022 and PRIN 2022-SAPPHIRE(CUP 2022MWK7Y7)funded by European Union-Next Generation EU,the“Tecnopolo per la medicina di precisione”(TecnoMed Puglia)-Regione Puglia:DGR n.2117 of 21/11/2018,CUP B84I18000540002)the Italian Ministry of Research(MUR)in the framework of the National Recovery and Resilience Plan(NRRP),“NFFA-DI”Grant(CUP B53C22004310006)“I-PHOQS”Grant(CUP B53C22001750006)under the complementary actions to the NRRP funded by NextGenerationEU,“Fit4MedRob”Grant(PNC0000007,CUP B53C22006960001).
文摘Understanding the interaction mechanisms of engineered nanomaterials(ENMs)with plant membranes is crucial for their effective use in various applications.While passive transport of smaller ENMs is well-documented,the mechanisms underlying active transport of larger ENMs remain poorly understood.This study systematically investigates the active transport and subcellular distribution of ENMs(100-1000 nm)within protoplasts using optical ratiometric silica pH sensors for localization.Highly monodispersed ratiometric pH sensors,based on silica particles modified with fluorescein-5-isothiocyanate(FITC)and cyanine3 NHS ester(CY3)dyes,were employed to elucidate internalization mechanisms.Protoplasts from Nicotiana tabacum L.leaves successfully internalized the sensors.3D segmentation of protoplasts revealed distinct pH gradients,indicating vacuole accumulation.Colocalization analysis and cellular compartments staining further confirmed sensor distribution.High-throughput imaging flow cytometry showed efficient internalization rates,which decreased after cell wall regeneration.Notably,inhibition experiments with the salicylic acid(SA)and Tyrphostin A23(TyrA23)inhibitors confirmed clathrin-mediated endocytosis in particle uptake.This study establishes rational design principles for controlling active ENM uptake and subcellular localization via optical pH sensing in protoplasts.The findings enhance our understanding of plant cell trafficking mechanisms and hold promise for targeted delivery and applications in plant biology research.
基金funding from the Italian Ministry for Education,University,and Research(MIUR)within the program“Departments of Excellence”2018-2022(DII-UNITN)from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement no.101008041.the NIH(P41EB027062)for support of this work.
文摘The shortage of tissues and organs for transplantation is an urgent clinical concern.In situ 3D printing is an advanced 3D printing technique aimed at printing the new tissue or organ directly in the patient.The ink for this process is central to the outcomes,and must meet specific requirements such as rapid gelation,shape integrity,stability over time,and adhesion to surrounding healthy tissues.Among natural materials,silk fibroin exhibits fascinating properties that have made it widely studied in tissue engineering and regenerative medicine.However,further improvements in silk fibroin inks are needed to match the requirements for in situ 3D printing.In the present study,silk fibroin-based inks were developed for in situ applications by exploiting covalent crosslinking process consisting of a pre-photo-crosslinking prior to printing and in situ enzymatic crosslinking.Two different silk fibroin molecular weights were characterized and the synergistic effect of the covalent bonds with shear forces enhanced the shift in silk secondary structure towardβ-sheets,thus,rapid stabilization.These hydrogels exhibited good mechanical properties,stability over time,and resistance to enzymatic degradation over 14 days,with no significant changes over time in their secondary structure and swelling behavior.Additionally,adhesion to tissues in vitro was demonstrated.
基金supported by Pioneer Hi-Bred International Inc.,A DuPont Company
文摘Sorghum （Sorghum bicolor） has high levels of starch, sugar, and fiber and is one of the most important energy crops in the world. Insect damage is one of the challenges that impacts sorghum biomass production. There are at least 150 insect species that can infest sorghum varieties worldwide. These insects can complete several generations within a growing season, they target various parts of sorghum plants at devel- opmental stages, and they cause significant biomass losses. Genetic research has revealed the existence of resistant genetics in sorghum and insect tolerant sorghum varieties have been identified. Various control methods have been developed, yet more effective management is needed for increasing sorghum biomass production. Although there are no transgenic sorghum products on the market yet, biotechnology has been recognized as an important tool for controlling insect pests and increasing sorghum production.
文摘In 1986, Binnig et al. developed the first atomic force microscope (AFM). The AFM, unlike the scanning tunnelling microscope (STM), has no demands for electrical conductivity, so it has been used in science and technology more widely. In 1988, the AFM was improved, and the AFM employing laser beam deflection for force detection (laser-AFM) was developed. In 1990, laser-AFM got the atomic-resolution. Up till now, the AFM has developed into a very important technique for studying the surface.
文摘The principle of scanning probe microscopes (SPM) was lust described by J. A. O’Keefe in the 1960s. In 1982, the scanning tunnelling microscope (STM), the first supreme example of SPM family, was developed; for which Binnig and Rohrer received the 1986 Nobel Prize in Physics. Shortly after that, in 1986 Binnig together with Quate and Gerber introduced the first atomic force microscope (AFM). Unlike the STM, the AFM
