Elesclomol (ELC) is an anticancer drug inducing mitochondria cytotoxicity through reactive oxygen species.Here,for the first time,we encapsulate the poorly water soluble ELC in monoolein-based cubosomes stabilized wit...Elesclomol (ELC) is an anticancer drug inducing mitochondria cytotoxicity through reactive oxygen species.Here,for the first time,we encapsulate the poorly water soluble ELC in monoolein-based cubosomes stabilized with Pluronic F127.Cellular uptake and nanocarrier accumulation close to the mitochondria with sub-micrometer distance is identified via three-dimensional (3D) confocal microscopy and edge-to-edge compartment analysis.To monitor the therapeutic effect of the ELC nanocarrier,we apply for the first time,label-free time-lapse multi-photon fluorescence lifetime imaging microscopy (MP-FLIM) to track NAD(P)H cofactors with sub-cellular resolution on live cells exposed to an anticancer nanocarrier.Improved in vitro cytotoxicity is verified when loading the pre-complexed ELC with copper (ELC-Cu).Importantly,for equivalent copper concentration,cubosomes loaded with ELC-Cu show higher cytotoxicity compared to the free drug.The novel nanocarrier shows promising features for systemic ELC-Cu administration,and furthermore we establish the MP-FLIM technique for the assessment of anticancer drug delivery systems.展开更多
Film-based fluoresce nt sen sors have been adapted into portable devices for the vapor phase detecti on.The satisfactory sensing performs nee(e.g.,sen sitivity,selectivity,reusability)largely bene fits from fluorophores.
Recently, fluorescence technique becomes very useful. It can allow for addressing a fundamental problem of cellular mechanism besides characterizing the species inside the cell to facilitate the diagnostic and prognos...Recently, fluorescence technique becomes very useful. It can allow for addressing a fundamental problem of cellular mechanism besides characterizing the species inside the cell to facilitate the diagnostic and prognostic value. Manipulation with fluorescent dyes provides many possibilities for their use as tags, probes, and sensors. These types can be intrinsic or extrinsic to the cell. They can become not only silent observers, but also participants, modulators or disruptors of specific activities outline the biological functions can be successfully studied quantitatively and qualitatively with fluorescence techniques.展开更多
To date,sperm morphometric studies have assessed whole sperm populations without considering sperm function.The aim of this study was to evaluate the relati on ship of sperm membrane and acrosomal integrity with sperm...To date,sperm morphometric studies have assessed whole sperm populations without considering sperm function.The aim of this study was to evaluate the relati on ship of sperm membrane and acrosomal integrity with sperm morphometry in liquid semen samples collected from bulls.To this end,sperm morphometry was performed on cryopreserved semen samples from 16 bulls by a combination of fluorescent dyes,including Hoechst 33343,carboxyfluorescein diacetate,and propidium iodide.This allowed discrimination of different subpopulations on the basis of sperm membrane and acrosomal integrity and analysis of the morphometries of the sperm head,nucleus,and acrosome using a specific plug-in module created on ImageJ.Acrosomal integrity was related to sperm morphometry as the heads of spermatozoa with a damaged acrosome were significantly smaller than those with a normal acrosome(P<0.001).In the case of spermatozoa with an intact acrosome,those with a damaged plasma membrane had a larger sperm head and acrosome than spermatozoa with an intact plasma membrane(P<0.001).No significant differences in the sperm head size were observed between sperm subpopulations without an acrosome or in the nuclear sperm morphometry of the different subpopulations.There was a positive correlation between the sperm motility values of the samples and the morphometric parameters for in tact spermatozoa.These correlations were particularly strong for the morphometric parameters of the sperm acrosome.We conclude that there are clear differences in the sperm morphometry depending on the status of the sperm membrane and acrosome and this should be considered when performing this kind of analysis.展开更多
文摘Elesclomol (ELC) is an anticancer drug inducing mitochondria cytotoxicity through reactive oxygen species.Here,for the first time,we encapsulate the poorly water soluble ELC in monoolein-based cubosomes stabilized with Pluronic F127.Cellular uptake and nanocarrier accumulation close to the mitochondria with sub-micrometer distance is identified via three-dimensional (3D) confocal microscopy and edge-to-edge compartment analysis.To monitor the therapeutic effect of the ELC nanocarrier,we apply for the first time,label-free time-lapse multi-photon fluorescence lifetime imaging microscopy (MP-FLIM) to track NAD(P)H cofactors with sub-cellular resolution on live cells exposed to an anticancer nanocarrier.Improved in vitro cytotoxicity is verified when loading the pre-complexed ELC with copper (ELC-Cu).Importantly,for equivalent copper concentration,cubosomes loaded with ELC-Cu show higher cytotoxicity compared to the free drug.The novel nanocarrier shows promising features for systemic ELC-Cu administration,and furthermore we establish the MP-FLIM technique for the assessment of anticancer drug delivery systems.
基金Finan cial support is ack no wledged from the Nati onal Natural Science Foundation of China(Nos.21527802,21673133,22002082,and 21820102005)the Higher Education Discipline Innovation Project(111 Project,No.B14041)the Fundamen tai Research Funds for the Central Universities(No.GK202005002).
文摘Film-based fluoresce nt sen sors have been adapted into portable devices for the vapor phase detecti on.The satisfactory sensing performs nee(e.g.,sen sitivity,selectivity,reusability)largely bene fits from fluorophores.
文摘Recently, fluorescence technique becomes very useful. It can allow for addressing a fundamental problem of cellular mechanism besides characterizing the species inside the cell to facilitate the diagnostic and prognostic value. Manipulation with fluorescent dyes provides many possibilities for their use as tags, probes, and sensors. These types can be intrinsic or extrinsic to the cell. They can become not only silent observers, but also participants, modulators or disruptors of specific activities outline the biological functions can be successfully studied quantitatively and qualitatively with fluorescence techniques.
基金This work was supported by the Spanish Ministry of Economy and Finance(MINECO)(grant AGL2017-85030-R)the European Territorial Cooperation Operational Program-Spain,France,and Andorra Area 2014-20(Program DietaPYR2 EFA144/16)and the DGA-FSE(grant A07_17R).We would like to acknowledge the use of the Research Support Service-SAI,University of Zaragoza.
文摘To date,sperm morphometric studies have assessed whole sperm populations without considering sperm function.The aim of this study was to evaluate the relati on ship of sperm membrane and acrosomal integrity with sperm morphometry in liquid semen samples collected from bulls.To this end,sperm morphometry was performed on cryopreserved semen samples from 16 bulls by a combination of fluorescent dyes,including Hoechst 33343,carboxyfluorescein diacetate,and propidium iodide.This allowed discrimination of different subpopulations on the basis of sperm membrane and acrosomal integrity and analysis of the morphometries of the sperm head,nucleus,and acrosome using a specific plug-in module created on ImageJ.Acrosomal integrity was related to sperm morphometry as the heads of spermatozoa with a damaged acrosome were significantly smaller than those with a normal acrosome(P<0.001).In the case of spermatozoa with an intact acrosome,those with a damaged plasma membrane had a larger sperm head and acrosome than spermatozoa with an intact plasma membrane(P<0.001).No significant differences in the sperm head size were observed between sperm subpopulations without an acrosome or in the nuclear sperm morphometry of the different subpopulations.There was a positive correlation between the sperm motility values of the samples and the morphometric parameters for in tact spermatozoa.These correlations were particularly strong for the morphometric parameters of the sperm acrosome.We conclude that there are clear differences in the sperm morphometry depending on the status of the sperm membrane and acrosome and this should be considered when performing this kind of analysis.
