Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an effic...Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an efficient platform for screening antiviral compounds and evaluating vaccine efficacy.In this study,we utilized the H9N2 subtype IAV as the working model.An 11-amino-acid HiBiT tag,derived from NanoLuc luciferase,was incorporated into the flexible linker region of the NS1 protein.Subsequently,the recombinant HiBiT-tagged virus was rescued.The recombinant virus exhibited high genetic stability and similar virological characteristics to the parental virus,both in vitro and in vivo.Particularly importantly,the replication profile of the HiBiT-tagged virus can be easily measured using the Nano-Glo assay system,achieving an efficient screening platform.Based on this platform,we have developed assays with both convenience and efficiency for screening antiviral reagents,evaluating immunization efficacy,and measuring neutralizing antibodies.展开更多
In this paper, the Radial Strain (RS) and Strain Rate (SR) was calculated using tagged MRI (tMRI) data. Using tagged magnetic resonance imaging (tMRI), the left ventricle short axis of five healthy adults (three men a...In this paper, the Radial Strain (RS) and Strain Rate (SR) was calculated using tagged MRI (tMRI) data. Using tagged magnetic resonance imaging (tMRI), the left ventricle short axis of five healthy adults (three men and two women) and four healthy male rats was imaged during diastolic and systolic phases on the mid-ventricle level. The RS and radial SR of the left ventricle were calculated at the mid-ventricular level of the cardiac cycle. The peak RS for rat and human heart was found to be 46.8 ± 0.68 and 40.7 ± 1.44, respectively, and it occurred at 40% of the cardiac cycle for both human and rat hearts. The peak systolic and diastolic radial SR for human heart was 1.10 ± 0.08 s- 1 and - 1.78 ± 0.02 s- 1, respectively, while it was 4.25 ± 0.02 s- 1 and - 5.16 ± 0.23 s- 1, respectively for rat heart. The results show that tMRI data can be used to characterize the cardiac function during systolic and diastolic phases of the cardiac cycle, and as a result, it can be used to evaluate the cardiac motion by calculating its RS and radial SR at different locations of the cardiac wall during both diastolic and systolic phases. This study also approves the validity of the tagged MRI data to accurately describe the radial cardiac motion.展开更多
Among various functional genomics tools used to characterize genes in plants, transposonbased mutagenesis approaches offer great potential, especially in barley and wheat, which possess large genomes and in which gene...Among various functional genomics tools used to characterize genes in plants, transposonbased mutagenesis approaches offer great potential, especially in barley and wheat, which possess large genomes and in which genetic transformation is not routine. Two Ds transposon flanking sequences(TNPs), TNP-29(27.4 c M(centi Morgan)) and TNP-79(70.3 c M), were mapped in the vicinity of a malting quality QTL located on chromosome 4H of barley. Reactivation of the Ds transposon sequence from these TNP lines led to the identification of genes in the malting QTL regions. Several Ds(dissociation) lines were generated by crossing TNP-29 and TNP-79 with an Ac TPase(activator) expressing line(25-B), and F2 progenies were subsequently screened for Ds insertions at new locations. To further characterize these Ds mutants, we mapped the new Ds flanking sequences on a barley genetic map and found that 29% of Ds were located in regions associated with the malting QTL located on chromosome 4H and in close proximity to other important malting-associated QTL across the barley chromosome. Using a sequence based approach, a linkage map was generated that confirmed the position of Ds loci in the barley genome map. Locating these Ds loci on the barley map opens avenues to dissect important malting QTL for facilitating identification of candidate malting genes.展开更多
Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunopreci...Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunoprecipitatin (iChIP), to isolate specific genomic regions retaining molecular interaction in order to perform non-biased identification of interacting molecules in vivo. Here, we developed a second-generation tagged LexA DNA-binding domain, 3xFNLDD, for the iChIP analysis. 3xFNLDD consists of 3 x FLAG tags, a nuclear localization signal (NLS), the DNA-binding domain (DB) and the dimerization domain of the LexA protein. Expression of 3xFNLDD can be detected by immunoblot analysis as well as flowcytometry. We showed that iChIP using 3xFNLDD is able to consistently isolate more than 10% of input genomic DNA, several-fold more efficient compared to the first-generation tagged LexA DB. 3xFNLDD would be a useful tool to perform the iChIP analysis for locus-specific biochemical epigenetics.展开更多
Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal ...Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal isotope-tagged antibodies instead of fluorophores,these techniques overcome spectral overlap limitations and enable high-dimensional,compensation-free analysis of complex biological systems at single-cell resolution.The performance of CyTOF and IMC critically depends on advanced nanomaterials labeled with stable metal isotopes,which are essential for improving sensitivity and multiplexing capacity.This review systematically discusses the design principles,synthesis methods,and functionalization strategies of mass-tagged nanomaterials tailored for CyTOF(e.g.,cell suspension analysis)and IMC(e.g.,spatial proteomics of tissue sections).We highlight their impactful applications in biomedicine,including proteomics,immunology,oncology,and neuroscience,emphasizing their roles in disease diagnosis,targeted drug development,and singlecell analysis.Despite these advancements,challenges such as nanomaterial biocompatibility,clinical scalability,and artificial intelligence(AI)-driven design are discussed,providing a roadmap for future research in personalized medicine and theranostics.展开更多
The energy of tagged photons, which were provided from the internal photon tagging system of the Laboratory of Nuclear Science, Tohoku University, has been calibrated using the d（γ,π-pp） reaction. Charged pions an...The energy of tagged photons, which were provided from the internal photon tagging system of the Laboratory of Nuclear Science, Tohoku University, has been calibrated using the d（γ,π-pp） reaction. Charged pions and protons in the final state were detected with the Neutral Kaon Spectrometer （NKS2）. Photon energies were obtained from the reaction of d（γ,π-pp）. The derived photon energy was consistent with the design of the tagger system and the previous measurement using electron-positron pair production. The consistency demonstrates the performance of NKS2 and the capability of the photon energy calibration using d（γ,π-pp）.展开更多
Stirring-exclusion processes are exclusion processes with particles being stirred. We investigate a tagged particle among a Bernoulli product environment measure on the lattice Zd. We show the strong law of large numb...Stirring-exclusion processes are exclusion processes with particles being stirred. We investigate a tagged particle among a Bernoulli product environment measure on the lattice Zd. We show the strong law of large numbers and the central limit theorem for the tagged particle. The proof of the central limit theorem is based on the method of martingale decomposition with a sector condition.展开更多
We investigate a tagged particle in the exclusion processes on {1,..., N }×Zd, with different densities in different levels {k} × Zd, ? k. Ignoring the level the tagged particle lying in, we only concern its...We investigate a tagged particle in the exclusion processes on {1,..., N }×Zd, with different densities in different levels {k} × Zd, ? k. Ignoring the level the tagged particle lying in, we only concern its position in Zd,denoted by Xt. Note that the whole space is not homogeneous. We define the environment process viewed from the tagged particle, of which Xt can be expressed as a functional. It is called the tagged particle process. We show the ergodicity of the tagged particle process, then prove the strong law of large numbers. Furthermore, we show the central limit theorem of Xt provided the zero-mean condition.展开更多
Virus tagged with greenfluorescent protein(GFP)contributes to the visualization and study of the virus in living cells.However,the hepatitis B virus(HBV)particle,which is a compact virion with limited internal space,ca...Virus tagged with greenfluorescent protein(GFP)contributes to the visualization and study of the virus in living cells.However,the hepatitis B virus(HBV)particle,which is a compact virion with limited internal space,cannot be incorporated with GFP tag as a large fragment.It was recently reported that protein genetically inserted with a smaller size tetracysteine(TC)tag could be specially labeled by a biarsenicalfluorescent dye in living cells.In this study,we constructed a recombinant HBV vector encoding TC-tagged core protein for biarsenical labeling of HBV virion.TC tag was genetically inserted near the immunodominant c/e1 site of HBV core protein by mutagenesis.Western blot and enzyme-linked immu-nosorbent assay(ELISA)analysis showed that the TC-tagged core protein,hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)could be expressed in cells transfected with the recombinant HBV vector,which is similar to the cells transfected with wild-type HBV vector.Reverse transcription-polymerase chain reaction(RT-PCR)and Southern blot analysis showed that HBV virion formation was affected by the genetic insertion of TC tag into core protein in some degree,but cells transfected with the HBV vector could still produce HBV virions incorporated with TC-tagged core proteins.Taken together,the recombinant HBV vector can serve as a useful tool to produce HBV virions incorporated with TC-tagged core proteins to befluorescently labeled by biarsencial dye for visualizing and studying HBV in living cells.展开更多
The mechanisms of mushroom polysaccharides on immune functions and lipid metabolism of aged mammals have not been fully elucidated.In the present study,after assessing the impacts of one type of Lentinula edodes-deriv...The mechanisms of mushroom polysaccharides on immune functions and lipid metabolism of aged mammals have not been fully elucidated.In the present study,after assessing the impacts of one type of Lentinula edodes-derived polysaccharides,named L2,on immune functions and blood lipid profiles,isobaric tags for relative and absolute quantification(iTRAQ)-based proteomic profiling of the small intestinal tissues from aged mice treated with L2 was performed.L2 reversed immune function declines and modulated the lipid metabolism of aged mice evidenced by increased levels of serum TC,HDL-C,and LDL-C,and reduced levels of serum TG.Moreover,a total of 95 differentially regulated proteins(DRPs) were identified,of which75 were up-regulated and 20 were down-regulated.Most of the DRPs were involved in intracellular and extracellular structure organization,and cellular and metabolic regulation.Particularly,approximately 16 and 9 DRPs participated in the regulation of immune functions and lipid metabolism,respectively.Furthermore,protein-protein interaction analysis highlighted that cadherin-1,plectin,cadherin-17,Ras GTPase-activating-like protein IQGAP2,and ezrin might be key proteins in response to L2 treatment.These findings provide new insights into the biological mechanisms of mushroom polysaccharides in anti-aging from a proteomic perspective.展开更多
基金the Animal Ethics Committee of the Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences(SYXK-2020-0010).
文摘Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an efficient platform for screening antiviral compounds and evaluating vaccine efficacy.In this study,we utilized the H9N2 subtype IAV as the working model.An 11-amino-acid HiBiT tag,derived from NanoLuc luciferase,was incorporated into the flexible linker region of the NS1 protein.Subsequently,the recombinant HiBiT-tagged virus was rescued.The recombinant virus exhibited high genetic stability and similar virological characteristics to the parental virus,both in vitro and in vivo.Particularly importantly,the replication profile of the HiBiT-tagged virus can be easily measured using the Nano-Glo assay system,achieving an efficient screening platform.Based on this platform,we have developed assays with both convenience and efficiency for screening antiviral reagents,evaluating immunization efficacy,and measuring neutralizing antibodies.
文摘In this paper, the Radial Strain (RS) and Strain Rate (SR) was calculated using tagged MRI (tMRI) data. Using tagged magnetic resonance imaging (tMRI), the left ventricle short axis of five healthy adults (three men and two women) and four healthy male rats was imaged during diastolic and systolic phases on the mid-ventricle level. The RS and radial SR of the left ventricle were calculated at the mid-ventricular level of the cardiac cycle. The peak RS for rat and human heart was found to be 46.8 ± 0.68 and 40.7 ± 1.44, respectively, and it occurred at 40% of the cardiac cycle for both human and rat hearts. The peak systolic and diastolic radial SR for human heart was 1.10 ± 0.08 s- 1 and - 1.78 ± 0.02 s- 1, respectively, while it was 4.25 ± 0.02 s- 1 and - 5.16 ± 0.23 s- 1, respectively for rat heart. The results show that tMRI data can be used to characterize the cardiac function during systolic and diastolic phases of the cardiac cycle, and as a result, it can be used to evaluate the cardiac motion by calculating its RS and radial SR at different locations of the cardiac wall during both diastolic and systolic phases. This study also approves the validity of the tagged MRI data to accurately describe the radial cardiac motion.
基金Funding for this project was provided by Barley Malting and Brewing Research Institute (grant number: 217248)
文摘Among various functional genomics tools used to characterize genes in plants, transposonbased mutagenesis approaches offer great potential, especially in barley and wheat, which possess large genomes and in which genetic transformation is not routine. Two Ds transposon flanking sequences(TNPs), TNP-29(27.4 c M(centi Morgan)) and TNP-79(70.3 c M), were mapped in the vicinity of a malting quality QTL located on chromosome 4H of barley. Reactivation of the Ds transposon sequence from these TNP lines led to the identification of genes in the malting QTL regions. Several Ds(dissociation) lines were generated by crossing TNP-29 and TNP-79 with an Ac TPase(activator) expressing line(25-B), and F2 progenies were subsequently screened for Ds insertions at new locations. To further characterize these Ds mutants, we mapped the new Ds flanking sequences on a barley genetic map and found that 29% of Ds were located in regions associated with the malting QTL located on chromosome 4H and in close proximity to other important malting-associated QTL across the barley chromosome. Using a sequence based approach, a linkage map was generated that confirmed the position of Ds loci in the barley genome map. Locating these Ds loci on the barley map opens avenues to dissect important malting QTL for facilitating identification of candidate malting genes.
文摘Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunoprecipitatin (iChIP), to isolate specific genomic regions retaining molecular interaction in order to perform non-biased identification of interacting molecules in vivo. Here, we developed a second-generation tagged LexA DNA-binding domain, 3xFNLDD, for the iChIP analysis. 3xFNLDD consists of 3 x FLAG tags, a nuclear localization signal (NLS), the DNA-binding domain (DB) and the dimerization domain of the LexA protein. Expression of 3xFNLDD can be detected by immunoblot analysis as well as flowcytometry. We showed that iChIP using 3xFNLDD is able to consistently isolate more than 10% of input genomic DNA, several-fold more efficient compared to the first-generation tagged LexA DB. 3xFNLDD would be a useful tool to perform the iChIP analysis for locus-specific biochemical epigenetics.
基金supported by National Natural Science Foundation of China(T2122002,82361148715,22077079,82204104)National Key R&D Program of China(2022YFC2601700,2022YFF0710202 and 2022YFA1104200)+4 种基金Princess Nourah bint Abdulrahman University Researchers Supporting Project number(PNURSP2024R122)Shanghai Municipal Science and Technology Projects(22Z510202478)Shanghai Municipal Education Commission Projects(21SG10,ZXWH1082101)Shanghai Jiao Tong University Projects(YG2021ZD19)Shanghai University of Medicine&Health Sciences Project(AMSCP-24-07-01).
文摘Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal isotope-tagged antibodies instead of fluorophores,these techniques overcome spectral overlap limitations and enable high-dimensional,compensation-free analysis of complex biological systems at single-cell resolution.The performance of CyTOF and IMC critically depends on advanced nanomaterials labeled with stable metal isotopes,which are essential for improving sensitivity and multiplexing capacity.This review systematically discusses the design principles,synthesis methods,and functionalization strategies of mass-tagged nanomaterials tailored for CyTOF(e.g.,cell suspension analysis)and IMC(e.g.,spatial proteomics of tissue sections).We highlight their impactful applications in biomedicine,including proteomics,immunology,oncology,and neuroscience,emphasizing their roles in disease diagnosis,targeted drug development,and singlecell analysis.Despite these advancements,challenges such as nanomaterial biocompatibility,clinical scalability,and artificial intelligence(AI)-driven design are discussed,providing a roadmap for future research in personalized medicine and theranostics.
基金Supported by a Grant-in-Aid (16GS0201) for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan
文摘The energy of tagged photons, which were provided from the internal photon tagging system of the Laboratory of Nuclear Science, Tohoku University, has been calibrated using the d（γ,π-pp） reaction. Charged pions and protons in the final state were detected with the Neutral Kaon Spectrometer （NKS2）. Photon energies were obtained from the reaction of d（γ,π-pp）. The derived photon energy was consistent with the design of the tagger system and the previous measurement using electron-positron pair production. The consistency demonstrates the performance of NKS2 and the capability of the photon energy calibration using d（γ,π-pp）.
文摘Stirring-exclusion processes are exclusion processes with particles being stirred. We investigate a tagged particle among a Bernoulli product environment measure on the lattice Zd. We show the strong law of large numbers and the central limit theorem for the tagged particle. The proof of the central limit theorem is based on the method of martingale decomposition with a sector condition.
基金supported by National Natural Science Foundation of China(Grant No.11371040)
文摘We investigate a tagged particle in the exclusion processes on {1,..., N }×Zd, with different densities in different levels {k} × Zd, ? k. Ignoring the level the tagged particle lying in, we only concern its position in Zd,denoted by Xt. Note that the whole space is not homogeneous. We define the environment process viewed from the tagged particle, of which Xt can be expressed as a functional. It is called the tagged particle process. We show the ergodicity of the tagged particle process, then prove the strong law of large numbers. Furthermore, we show the central limit theorem of Xt provided the zero-mean condition.
基金supported in part by the National Natural Science Foundation of China(Grant Nos.30872237 and 30600277)National Key Basic Research Program of China(973 Program,No.2007CB512900)Doctoral Fund of Ministry of Education of China(No.20070487007).
文摘Virus tagged with greenfluorescent protein(GFP)contributes to the visualization and study of the virus in living cells.However,the hepatitis B virus(HBV)particle,which is a compact virion with limited internal space,cannot be incorporated with GFP tag as a large fragment.It was recently reported that protein genetically inserted with a smaller size tetracysteine(TC)tag could be specially labeled by a biarsenicalfluorescent dye in living cells.In this study,we constructed a recombinant HBV vector encoding TC-tagged core protein for biarsenical labeling of HBV virion.TC tag was genetically inserted near the immunodominant c/e1 site of HBV core protein by mutagenesis.Western blot and enzyme-linked immu-nosorbent assay(ELISA)analysis showed that the TC-tagged core protein,hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)could be expressed in cells transfected with the recombinant HBV vector,which is similar to the cells transfected with wild-type HBV vector.Reverse transcription-polymerase chain reaction(RT-PCR)and Southern blot analysis showed that HBV virion formation was affected by the genetic insertion of TC tag into core protein in some degree,but cells transfected with the HBV vector could still produce HBV virions incorporated with TC-tagged core proteins.Taken together,the recombinant HBV vector can serve as a useful tool to produce HBV virions incorporated with TC-tagged core proteins to befluorescently labeled by biarsencial dye for visualizing and studying HBV in living cells.
基金supported by the Key-Area Research and Development Program of Guangdong Province (2021B0707060001)the Program for Scientific Research Start-Up Funds of Guangdong Ocean UniversityChina Postdoctoral Science Foundation (2016T90787)。
文摘The mechanisms of mushroom polysaccharides on immune functions and lipid metabolism of aged mammals have not been fully elucidated.In the present study,after assessing the impacts of one type of Lentinula edodes-derived polysaccharides,named L2,on immune functions and blood lipid profiles,isobaric tags for relative and absolute quantification(iTRAQ)-based proteomic profiling of the small intestinal tissues from aged mice treated with L2 was performed.L2 reversed immune function declines and modulated the lipid metabolism of aged mice evidenced by increased levels of serum TC,HDL-C,and LDL-C,and reduced levels of serum TG.Moreover,a total of 95 differentially regulated proteins(DRPs) were identified,of which75 were up-regulated and 20 were down-regulated.Most of the DRPs were involved in intracellular and extracellular structure organization,and cellular and metabolic regulation.Particularly,approximately 16 and 9 DRPs participated in the regulation of immune functions and lipid metabolism,respectively.Furthermore,protein-protein interaction analysis highlighted that cadherin-1,plectin,cadherin-17,Ras GTPase-activating-like protein IQGAP2,and ezrin might be key proteins in response to L2 treatment.These findings provide new insights into the biological mechanisms of mushroom polysaccharides in anti-aging from a proteomic perspective.
