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Dual Targeting to Mitochondria and Chloroplasts： Characterization of Thr-tRNA Synthetase Targeting Peptide

Molecular Plant 2009年第6期2卷 1298-1309页

作者： Anna-Karin Berglund Erika Spanning Henrik Biverstahl Gianluca Maddalo Christian Tellgren-Roth lena Maler Elzbieta Glaser Department of Biochemistry and Biophysics Arrhenius Laboratories for Natural Science Stockholm University SF-10691 Stockholm Sweden Department of Genetics and Pathology Rudbecklaboratoriet Uppsala University SE-751 85 Uppsala Sweden

There is a group of proteins that are encoded by a single gene, expressed as a single precursor protein and dually targeted to both mitochondria and chloroplasts using an ambiguous targeting peptide. Sequence analysis of 43 dual targeted proteins in comparison with 385 mitochondrial proteins and 567 chloroplast proteins ofArabidopsis thaliana revealed an overall significant increase in phenylalanines, leucines, and serines and a decrease in acidic amino acids and glycine in dual targeting peptides （dTPs）. The N-terminal portion of dTPs has significantly more serines than mTPs. The number of arginines is similar to those in mTPs, but almost twice as high as those in cTPs. We have investigated targeting determinants of the dual targeting peptide of Thr-tRNA synthetase （ThrRS-dTP） studying organellar import of N- and C-terminal deletion constructs of ThrRS-dTP coupled to GFR These results show that the 23 amino acid long N-terminal portion of ThrRS-dTP is crucial but not sufficient for the organellar import. The C-terminal deletions revealed that the shortest peptide that was capable of conferring dual targeting was 60 amino acids long. We have purified the ThrRS- dTP（2-60） to homogeneity after its expression as a fusion construct with GST followed by CNBr cleavage and ion exchange chromatography. The purified ThrRS-dTP（2-60） inhibited import of pF1β into mitochondria and of pSSU into chloroplasts at μM concentrations showing that dual and organelle-specific proteins use the same organellar import pathways. Furthermore, the CD spectra of ThrRS-dTP（2-60） indicated that the peptide has the propensity for forming α-helical structure in membrane mimetic environments; however, the membrane charge was not important for the amount of induced helical structure. This is the first study in which a dual targeting peptide has been purified and investigated by biochemical and biophysical means.

关键词： Dual targeting mitochondria chloroplast targeting peptide aminoacyl-tRNA synthetase

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Single-cell trajectories of melanoma cell resistance to targeted treatment

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Cancer Biology & Medicine 2022年第1期19卷 56-73页

作者： Maria Schmidt lena Sünke Mortensen Henry Loeffler-Wirth Corinna Kosnopfel Knut Krohn Hans Binder Manfred Kunz Interdisciplinary Centre for Bioinformatics University of LeipzigLeipzig 04107Germany Department of Dermatology Venereology and AllergologyUniversity of WürzburgWürzburg 97074Germany Core Unit DNA Technologies Medical FacultyUniversity of LeipzigLeipzig 04103Germany Department of Dermatology Venereology and AllergologyUniversity of Leipzig Medical CenterLeipzig 04103Germany

Objective:Cellular heterogeneity is regarded as a major factor affecting treatment response and resistance in malignant *** developments in single-cell sequencing technology have provided deeper insights into these ***:Here,we analyzed a BRAFV600 E-mutant melanoma cell line by single-cell RNA-seq under various conditions:cells sensitive to BRAF inhibition with BRAF inhibitor vemurafenib and cells resistant to BRAF inhibition with vemurafenib alone or vemurafenib in combination with the MEK1/2 inhibitors cobimetinib or *** reduction by t-distributed stochastic neighbor embedding and self-organizing maps identified distinct trajectories of resistance development clearly separating the 4 treatment conditions in cell and gene state ***:Trajectories associated with resistance to single-agent treatment involved cell cycle,extracellular matrix,and de-differentiation *** contrast,shifts detected in double-resistant cells primarily affected translation and mitogen-activated protein kinase pathway reactivation,with a small subpopulation showing markers of *** findings were validated in pseudotime analyses and RNA velocity ***:The single-cell transcriptomic analyses reported here employed a spectrum of bioinformatics methods to identify mechanisms of melanoma resistance to single-and double-agent *** study deepens our understanding of treatmentinduced cellular reprogramming and plasticity in melanoma cells and identifies targets of potential relevance to the management of treatment resistance.

关键词： Melanoma single-cell transcriptome sequencing treatment response

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THE SMUGGLING ISSUE IN GOLD WE LUST

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城市漫步（GBA版） 2014年第6期 36-45页

作者： lena GIDWANI 不详

In the mid-sixth century;two monks snuck silkworm larvae out of China in bamboo canes,their act of treason starting the silk industry of the Byzantine *** gave them a silk monopoly in Europe,the foundation for the Byzantine economy until its demise some 650 years later In the 1520s,South Chinas coast was a hotbed of illicit com-merce.

关键词： starting silk foundation

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Orchestration of antiviral responses within the infected central nervous system

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Cellular & Molecular Immunology 2024年第9期21卷 943-958页

作者： Andreas Pavlou Felix Mulenge Olivia Luise Gern lena Mareike Busker Elisabeth Greimel Inken Waltl Ulrich Kalinke Institute for Experimental Infection Research TWINCORECentre for Experimental and Clinical Infection Researcha joint venture between the Helmholtz Centre for Infection Research and the Hannover Medical School30625HannoverGermany Department of Pathology University of Veterinary Medicine HannoverFoundation30559HannoverGermany Cluster of Excellence RESIST(EXC 2155) Hannover Medical School30625HannoverGermany

Many newly emerging and re-emerging viruses have neuroinvasive potential,underscoring viral encephalitis as a global research *** entry of the virus into the CNS,severe neurological life-threatening conditions may manifest that are associated with high morbidity and *** currently available therapeutic arsenal against viral encephalitis is rather limited,emphasizing the need to better understand the conditions of local antiviral immunity within the infected *** this review,we discuss new insights into the pathophysiology of viral encephalitis,with a focus on myeloid cells and CD8^(+)T cells,which critically contribute to protection against viral CNS *** illuminating the prerequisites of myeloid and T cell activation,discussing new discoveries regarding their transcriptional signatures,and dissecting the mechanisms of their recruitment to sites of viral replication within the CNS,we aim to further delineate the complexity of antiviral responses within the infected ***,we summarize the current knowledge in the field of virus infection and neurodegeneration and discuss the potential links of some neurotropic viruses with certain pathological hallmarks observed in neurodegeneration.

关键词： Virus infection Virus control within the brain T cell recruitment to the brain Microglia activation

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Haplotype resolved chromosome level genome assembly of Citrus australis reveals disease resistance and other citrus specific genes

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Horticulture Research 2023年第5期10卷 193-206页

作者： Upuli Nakandala Ardashir Kharabian Masouleh Malcolm WSmith Agnelo Furtado Patrick Mason lena Constantin Robert J.Henry Queensland Alliance for Agriculture and Food Innovation University of QueenslandBrisbane 4072Australia ARC Centre of Excellence for Plant Success in Nature and Agriculture University of QueenslandBrisbane 4072Australia Department of Agriculture and Fisheries Bundaberg Research StationBundabergQueensland 4670Australia

Recent advances in genome sequencing and assembly techniques have made it possible to achieve chromosome level reference genomes for *** few genomes have been anchored at the chromosome level and/or are haplotype phased,with the available genomes of varying accuracy and *** now report a phased high-quality chromosome level genome assembly for an Australian native citrus species;Citrus australis(round lime)using highly accurate PacBio HiFi long reads,complemented with Hi-C *** with Hi-C integrated assembly resulted in a 331 Mb genome of *** with two haplotypes of nine pseudochromosomes with an N50 of 36.3 Mb and 98.8% genome assembly completeness(BUSCO).Repeat analysis showed that more than 50% of the genome contained interspersed *** them,LTR elements were the predominant type(21.0%),of which LTR Gypsy(9.8%)and LTR copia(7.7%)elements were the most abundant repeats.A total of 29464 genes and 32009 transcripts were identified in the *** these,28222 CDS(25753 genes)had BLAST hits and 21401 CDS(75.8%)were annotated with at least one GO *** specific genes for antimicrobial peptides,defense,volatile compounds and acidity regulation were *** synteny analysis showed conserved regions between the two haplotypes with some structural variations in Chromosomes 2,4,7 and *** chromosome scale,and haplotype resolved *** genome will facilitate the study of important genes for citrus breeding and will also allow the enhanced definition of the evolutionary relationships between wild and domesticated citrus species.

关键词： citrus Citrus breeding

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Effect of processing and accelerated storage on the volatile composition and sensory profile of a tomato soup

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Food Quality and Safety 2022年第2期6卷 341-352页

作者： lena C.Strohla Krishnachandra S.Hidangmayum Sandra S.Waehrens Vibeke Orlien Mikael A.Petersen Department of Food Science University of Copenhagen1958 Frederiksberg CDenmark

The volatile and sensory profile of pasteurized(95℃,18 min)and sterilized(120℃,14 min)tomato soup and its changes at accelerated storage conditions(37℃and 42℃)were identified,characterized,and correlated by predictive *** led to more profound changes over storage compared to pasteurisation,showing enhanced levels of‘dark'odour,‘burnt'flavour,and‘complexity',while pasteurized soup had more pronounced‘green'odour,‘vegetables'flavour,‘harmony',and‘freshness'.Over time,‘dark'and‘burnt'significantly increased for sterilized soups,whereas‘green'decreased significantly only for sterilized soup stored at 42℃.The changes of‘dark',‘green',‘off-flavour,‘burnt',‘har mony',and‘freshness'were associated with formation of furans(furfural and 2-acetyl-5-methylfuran),and decline of unsaturated aldehydes like E-2-octenal and E,E-2,*** compounds were identified to represent potential shelf-life markers,yet more studies on their odour activity in the tomato soup matrix are necessary to understand their contribution to the sensory *** construction of partial least squares models allowed the prediction of‘green',‘off-flavour',‘burnt',and‘freshness'based on selected *** models will be important tools in the process of predicting the end of shelf life,but need to be supplemented with consumer acceptability levels.

关键词： Volatile composition sensory profile accelerated storage tomato soup shelf life prediction partial least dquares regression

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Solriamfetol impurities:Synthesis,characterization,and analytical method(UPLC-UV)validation

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Journal of Pharmaceutical Analysis 2023年第4期13卷 403-411页

作者： Nafisah Al-Rifai Anas Alshishani Fouad Darras Ola Taha Shereen Abu-Jalloud lena Shaghlil Yousef Al-Ebini Pharmaceutical and Chemical Engineering Department School of Medical SciencesGerman Jordanian UniversityAmmanJordan Faculty of Pharmacy Zarqa UniversityZarqaJordan Research and Development Department Resonance Research LabAmmanJordan Department of Pharmaceutics College of PharmacyKing Khalid UniversityAbhaSaudi Arabia

Given that impurities may affect the quality and safety of drug products,impurity identification and profiling is an integral part of drug quality control and is particularly important for newly developed medications such as solriamfetol,which is used to treat excessive daytime *** the highperformance liquid chromatography analysis of commercial solriamfetol has revealed the presence of several impurities,their synthesis,structure elucidation,and chromatographic determination have not been reported *** bridge this gap,we herein identified,synthesized,and isolated eight processrelated solriamfetol impurities,characterized them using spectroscopic and chromatographic techniques,and proposed plausible mechanisms of their ***,we developed and validated a prompt impurity analysis method based on ultrahigh-performance liquid chromatography with UV detection,revealing that its selectivity,linearity,accuracy,precision,and quantitation limit meet the acceptance criteria of method validation stipulated by the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human ***,the developed method was concluded to be suitable for the routine analysis of solriamfetol substances.

关键词： Solriamfetol Impurity analysis Impurity synthesis UPLC Method validation

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Apoplastic barriers of Populus×canescens roots in reaction to different cultivation conditions and abiotic stress treatments

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Stress Biology 2023年第1期3卷 269-282页

作者： Paul Grunhofer Ines Heimerich lena Herzig Svenja Pohl Lukas Schreiber Department of Ecophysiology Institute of Cellular and Molecular BotanyUniversity of BonnKirschallee 153115 BonnGermany

Populus is an important tree genus frequently cultivated for economical ***,the high sensitivity of poplars towards water deficit,drought,and salt accumulation significantly affects plant productivity and limits biomass *** cultivation and abiotic stress conditions have been described to significantly induce the formation of apoplastic barriers(Casparian bands and suberin lamellae)in roots of different monocotyledonous crop ***,this study aimed to investigate to which degree the roots of the dicotyledonous gray poplar(Populus×canescens)react to a set of selected cultivation conditions(hydroponics,aeroponics,or soil)and abiotic stress treatments(abscisic acid,oxygen deficiency)because a differing stress response could potentially help in explaining the observed higher stress *** apoplastic barriers of poplar roots cultivated in different environments were analyzed by means of histochemistry and gas chromatography and compared to the available literature on monocotyledonous crop ***,dicotyledonous poplar roots showed only a remarkably low induction or enhancement of apoplastic barriers in response to the different cultivation conditions and abiotic stress *** genetic optimization(e.g.,overexpression of biosynthesis key genes)of the apoplastic barrier development in poplar roots might result in more stress-tolerant cultivars in the future.

关键词： Hydroponics Aeroponics Soil Abscisic acid(ABA) Oxygen deficiency Poplar root suberin

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Promoting the production of challenging proteins via induced expression in CHO cells and modified cell-free lysates harboring T7 RNA polymerase and mutant eIF2α

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Synthetic and Systems Biotechnology 2024年第3期9卷 416-424页

作者： Jeffrey L.Schloßhauer lena Tholen Alexander Korner Stefan Kubick Sofia Chatzopoulou Anja Honow Anne Zemella Fraunhofer Project Group PZ-Syn of the Fraunhofer Institute for Cell Therapy and Immunology-IZI Branch Bioanalytics and Bioprocesses-IZI-BBAm MühlenbergPotsdamGermany Fraunhofer Institute for Cell Therapy and Immunology-IZI Branch Bioanalytics and Bioprocesses-IZI-BBAm MühlenbergPotsdamGermany Faculty of Health Sciences Joint Faculty of the Brandenburg University of Technology Cottbus-SenftenbergThe Brandenburg Medical School Theodor FontaneUniversity of PotsdamPotsdamGermany Institute for Chemistry and Biochemistry Laboratory of Protein BiochemistryFreie Universitat BerlinThielallee 6314195BerlinGermany Institute of Biotechnology Technische Universit¨at BerlinStraße des 17.Juni 13510623BerlinGermany New/era/mabs GmbH August-Bebel-Str.8914482PotsdamGermany

Chinese hamster ovary(CHO)cells are crucial in biopharmaceutical production due to their scalability and capacity for human-like post-translational ***,toxic proteins and membrane proteins are often difficult-to-express in living ***,cell-free protein synthesis can be *** study explores innovative strategies for enhancing the production of challenging proteins through the modification of CHO cells by investigating both,cell-based and cell-free approaches.A major result in our study involves the integration of a mutant eIF2 translation initiation factor and T7 RNA polymerase into CHO cell lysates for cell-free protein *** resulted in elevated yields,while eliminating the necessity for exogenous additions during cell-free production,thereby substantially enhancing ***,we explore the potential of the Rosa26 genomic site for the integration of T7 RNA polymerase and cell-based tetracycline-controlled protein *** findings provide promising advancements in bioproduction technologies,offering flexibility to switch between cell-free and cell-based protein production as needed.

关键词： Inducible expression CHO cells Cell-free protein synthesis CRISPR T7 RNA polymerase eIF2 Rosa26

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Sex differences and effects of oestrogen in rat gastric mucosal defence

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World Journal of Gastroenterology 2017年第3期23卷 426-436页

作者： Richard Shore Hakan Bjorne Yoko Omoto Anna Siemiatkowska Jan-Ake Gustafsson Mats Lindblad lena Holm Department of Clinical Sciences Intervention and Technology(CLINTEC)Karolinska InstitutetS-14186 StockholmSweden Function Perioperative Medicine and Intensive Care Karolinska University HospitalS-17176 StockholmSweden Department of Physiology and Pharmacology Karolinska InstitutetS-17177 StockholmSweden Department of Endocrine and Breast Surgery Graduate School of Medical ScienceKyoto Prefectural University of Medicine 465 Kajii-choHirokoji AgaruKawaramachi-doriKamigyo-kuKyoto 602-0841Japan Department of Medical Cell Biology Uppsala UniversityS-75123 UppsalaSweden Center of Nuclear Receptors and Cell Signalling University of HoustonHoustonTX 77004United States Department of Clinical Sciences Intervention and Technology(CLINTEC)Karolinska InstitutetS-14186 StockholmSweden

AIM To evaluate sex differences and the effects of oestrogen administration in rat gastric mucosal *** Sex differences in gastric mucus thickness and accumulation rate, absolute gastric mucosal blood flow using microspheres, the integrity of the gastric mucosal epithelium in response to a chemical irritant and the effects of oestrogen administration on relative gastric mucosal blood flow in an acute setting was assessed in an in vivo rat experimental model. Subsequently, sex differences in the distribution of oestrogen receptors and calcitonin gene related peptide in the gastric mucosa of animals exposed to oestrogen in the above experiments was evaluated using *** The absolute blood flow in the GI-tract was generally higher in males, but only significantly different in the corpus part of the stomach (1.12 ± 0.12 m L/min·g in males and 0.51 ± 0.03 m L/min·g in females) (P = 0.002). After removal of the loosely adherent mucus layer the thickness of the firmly adherent mucus layer in males and females was 79 ± 1 μm and 80 ± 3 μm respectively. After 60 min the mucus thickness increased to 113 ± 3 μm in males and 121 ± 3 μm in females with no statistically significant difference seen between the sexes. Following oestrogen administration(0.1 followed by 1 μg/kg·min), mean blood flow in the gastric mucosa decreased by 31% [68 ± 13 perfusion units (PFU)] in males which was significantly different compared to baseline(P = 0.02). In females however, mean blood flow remained largely unchanged with a 4% (5 ± 33 PFU) reduction. The permeability of the gastric mucosa increased to a higher level in females than in males (P = 0.01) after taurocholate challenge. However, the calculated mean clearance increase did not significantly differ between the sexes [0.1 ± 0.04 to 1.1 ± 0.1 m L/min·100 g in males and 0.4 ± 0.3 to 2.1 ± 0.3 m L/min·100 g in females(P = 0.065)]. There were no significant differences between 17β-Estradiol treated males (me

关键词： Sex differences Gastric mucosal defence Blood flow Oestrogen Gastric physiology Mucus

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