المؤلفون: Ann-Kathrin Kissmann, Barbara Spellerberg, Pengfei Xu, Heinz Fabian Raber, Hu Xing, Daniel Mayer, Nicholas Bodenberger, Tanja Weil, Dennis Kubiczek, Uwe Knippschild, Deena D’souza, Frank Rosenau

المساهمون: European Union (EU), Horizon 2020

المصدر: International Journal of Molecular Sciences
Volume 22
Issue 19
International Journal of Molecular Sciences, Vol 22, Iss 10425, p 10425 (2021)

مصطلحات موضوعية: DDC 540 / Chemistry & allied sciences, 02 engineering and technology, Feces, DDC 570 / Life sciences, Biology (General), Spectroscopy, 0303 health sciences, biology, Chemistry, SELEX Aptamer Technique, General Medicine, Aptamers, Nucleotide, 021001 nanoscience & nanotechnology, 3. Good health, Computer Science Applications, ddc:540, 0210 nano-technology, Akkermansia muciniphila, Aptamer, QH301-705.5, aptamers, Computational biology, Article, Catalysis, Inorganic Chemistry, 03 medical and health sciences, Human gut, Alzheimer Disease, ddc:570, FluCell-SELEX, Humans, Molecule, Physical and Theoretical Chemistry, QD1-999, Molecular Biology, 030304 developmental biology, Oligonucleotide, Organic Chemistry, Akkermansia, biology.organism_classification, biosensors, Gastrointestinal Microbiome, Biosensors, Biosensor, Systematic evolution of ligands by exponential enrichment, Bacteria

الوصف: Based on their unique properties, oligonucleotide aptamers have been named a gift of biological chemistry to life science. We report the development of DNA aptamers as the first high-affinity binding molecules available for fast and rapid labeling of the human gut bacterium Akkermansia muciniphila with a certain impact on Alzheimer´s disease. Fast and reliable analyses of the composition of microbiomes is an emerging field in microbiology. We describe the molecular evolution and biochemical characterization of a specific aptamer library by a FluCell-SELEX and the characterization of specific molecules from the library by bioinformatics. The aptamer AKK13.1 exerted universal applicability in different analysis techniques in modern microbiology, including fluorimetry, confocal laser scanning microscopy and flow cytometry. It was also functional as a specific binding entity hybridized to anchor primers chemically coupled via acrydite-modification to the surface of a polyacrylamide-hydrogel, which can be prototypically used for the construction of affinity surfaces in sensor chips. Together, the performance and methodological flexibility of the aptamers presented here may open new routes not only to develop novel Akkermansia-specific assays for clinical microbiology and the analyses of human stool samples but may also be an excellent starting point for the construction of novel electronic biosensors.
publishedVersion

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a3015933ed409c5959ed7b5229585636Test

المؤلفون: Steffen Stenger, Jens Michaelis, Paul Walther, Reiner Noschka, Tanja Weil, Fanny Wondany, Gilbert Weidinger, Gönül Kizilsavas

المصدر: International Journal of Molecular Sciences, Vol 22, Iss 8392, p 8392 (2021)
International Journal of Molecular Sciences
Volume 22
Issue 16

مصطلحات موضوعية: Antigens, Differentiation, T-Lymphocyte, 0301 basic medicine, antimicrobial peptide, QH301-705.5, Human pathogen, Article, Catalysis, Microbiology, Inorganic Chemistry, Mycobacterium tuberculosis, 03 medical and health sciences, 0302 clinical medicine, Animals, Humans, human, Physical and Theoretical Chemistry, Granulysin, Biology (General), Cytotoxicity, Molecular Biology, Pathogen, QD1-999, Cells, Cultured, Zebrafish, Spectroscopy, biology, Chemistry, Organic Chemistry, General Medicine, biology.organism_classification, Antimicrobial, Computer Science Applications, macrophages, Cytolysis, 030104 developmental biology, tuberculosis, 030220 oncology & carcinogenesis, Host-Pathogen Interactions, Peptides, Intracellular

الوصف: Granulysin is an antimicrobial peptide (AMP) expressed by human T-lymphocytes and natural killer cells. Despite a remarkably broad antimicrobial spectrum, its implementation into clinical practice has been hampered by its large size and off-target effects. To circumvent these limitations, we synthesized a 29 amino acid fragment within the putative cytolytic site of Granulysin (termed “Gran1”). We evaluated the antimicrobial activity of Gran1 against the major human pathogen Mycobacterium tuberculosis (Mtb) and a panel of clinically relevant non-tuberculous mycobacteria which are notoriously difficult to treat. Gran1 efficiently inhibited the mycobacterial proliferation in the low micro molar range. Super-resolution fluorescence microscopy and scanning electron microscopy indicated that Gran1 interacts with the surface of Mtb, causing lethal distortions of the cell wall. Importantly, Gran1 showed no off-target effects (cytokine release, chemotaxis, cell death) in primary human cells or zebrafish embryos (cytotoxicity, developmental toxicity, neurotoxicity, cardiotoxicity). Gran1 was selectively internalized by macrophages, the major host cell of Mtb, and restricted the proliferation of the pathogen. Our results demonstrate that the hypothesis-driven design of AMPs is a powerful approach for the identification of small bioactive compounds with specific antimicrobial activity. Gran1 is a promising component for the design of AMP-containing nanoparticles with selective activity and favorable pharmacokinetics to be pushed forward into experimental in vivo models of infectious diseases, most notably tuberculosis.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4f0b2b05c96e6bf7fa1f8b3428c0f98aTest
https://www.mdpi.com/1422-0067/22/16/8392Test

المؤلفون: Valerie Amann, Ann-Kathrin Kissmann, Markus Krämer, Imke Krebs, Julio A. Perez-Erviti, Anselmo J. Otero-Gonzalez, Fidel Morales-Vicente, Armando Rodríguez, Ludger Ständker, Tanja Weil, Frank Rosenau

المصدر: Pharmaceutics

مصطلحات موضوعية: 03 medical and health sciences, 0302 clinical medicine, Pharmaceutical Science, 02 engineering and technology, 021001 nanoscience & nanotechnology, 0210 nano-technology, 030226 pharmacology & pharmacy, 3. Good health

الوصف: Antimicrobial peptides (AMPs) are an alternative group for the therapy of infectious diseases, with activity against a wide range of diverse pathogens. However, classical AMPs have significant side effects in human cells due to their unspecific pore formation in biomembranes. Nevertheless, AMPs are promising therapeutics and can be isolated from natural sources, which include sea and freshwater molluscs. The AMPs identified in these organisms show promising antimicrobial activities, as pathogens are mainly fought by innate defence mechanisms. An auspicious candidate among molluscs is the Cuban freshwater snail Pomacea poeyana, from which the peptides Pom-1 and Pom-2 have been isolated and studied. These studies revealed significant antimicrobial activities for both AMPs. Based on the activities determined, Pom-1 was used for further optimization. In order to meet the emerging requirements of improved anti-biofilm activity against naturally occurring Candida species, the six derivatives Pom-1A to F were developed and investigated. Analysis of the derivatives acting on the most abundant naturally occurring Candida yeast Candida albicans (C. albicans) revealed a strong anti-biofilm activity, especially induced by Pom-1 B, C, and D. Furthermore, a moderate decrease in the metabolic activity of planktonic yeast cells was observed.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4f5d0d39be34e236fed48b978ad9fbc2Test
https://doi.org/10.3390/pharmaceutics14020318Test

المؤلفون: Bikram Keshari Agrawalla, Andreas Riegger, Matthias P. Domogalla, Tao Wang, Xi Chen, Thilo Dörfler, Tanja Weil, Felix Boldt, Seah Ling Kuan, Markus Lamla, Jens Michaelis, Kerstin Steinbrink

المصدر: Bioconjugate Chemistry

مصطلحات موضوعية: 0301 basic medicine, Models, Molecular, Biomedical Engineering, Pharmaceutical Science, Bioengineering, 010402 general chemistry, 01 natural sciences, law.invention, Cell Line, Maleimides, 03 medical and health sciences, chemistry.chemical_compound, Mice, Bacterial Proteins, law, Animals, Humans, Reactivity (chemistry), Cysteine, Sulfhydryl Compounds, Sulfones, Maleimide, Peptide sequence, Dual labeling, Pharmacology, chemistry.chemical_classification, Staining and Labeling, Communication, Organic Chemistry, Disulfide bond, Proteins, Combinatorial chemistry, Recombinant Proteins, 0104 chemical sciences, Allyl Compounds, Luminescent Proteins, 030104 developmental biology, chemistry, Thiol, Recombinant DNA, Surface modification, Interleukin-2, Peptides, Biotechnology

الوصف: The attachment of two different functionalities in a site-selective fashion represents a great challenge in protein chemistry. We report site specific dual functionalizations of peptides and proteins capitalizing on reactivity differences of cysteines in their free (thiol) and protected, oxidized (disulfide) forms. The dual functionalization of interleukin 2 and EYFP proceeded with no loss of bioactivity in a stepwise fashion applying maleimide and disulfide rebridging allyl-sulfone groups. In order to ensure broader applicability of the functionalization strategy, a novel, short peptide sequence that introduces a disulfide bridge was designed and site-selective dual labeling in the presence of biogenic groups was successfully demonstrated.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2ef9ee7f463719fd66ae91cff542eb82Test
http://europepmc.org/articles/PMC6242188Test

المؤلفون: Karin Soller, Tanja Weil, Jose A. Cancelas, Novella Guidi, Johannes M. Weiss, Ludger Ständker, Gina Marka, Frank Kirchhoff, Hartmut Geiger, Karina Eiwen, Maria Carolina Florian, Mehmet Sacma

المصدر: The EMBO Journal
EMBO Journal

مصطلحات موضوعية: 0301 basic medicine, Aging, Myeloid, osteopontin, Stem cells, localization, alpha(9)beta(1), Mice, 0302 clinical medicine, DDC 570 / Life sciences, Osteopontin, requirements, stromal cells, 0303 health sciences, General Neuroscience, Stem Cells, Hematopoietic stem cell, hemic and immune systems, progenitor cells, Articles, Corrigenda, Maus, Cell biology, Haematopoiesis, niche, Phenotype, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Stem cell, Stromal cell, Biology, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, stomatognathic system, ddc:570, component, medicine, Animals, Regeneration, Humans, Progenitor cell, Molecular Biology, 030304 developmental biology, Nische, bone-marrow niche, General Immunology and Microbiology, Altern, Hematopoietic Stem Cells, microenvironment, Mice, Inbred C57BL, Lokalisation, Ageing, 030104 developmental biology, biology.protein, Mesenchymal stem cells, hematopoietic stem cell, Bone marrow, Cell Adhesion, Polarity & Cytoskeleton, Hematopoietic stem cells

الوصف: Upon aging, hematopoietic stem cells (HSCs) undergo changes in function and structure, including skewing to myeloid lineages, lower reconstitution potential and loss of protein polarity. While stem cell intrinsic mechanisms are known to contribute to HSC aging, little is known on whether age-related changes in the bone marrow niche regulate HSC aging. Upon aging, the expression of osteopontin (OPN) in the murine bone marrow stroma is reduced. Exposure of young HSCs to an OPN knockout niche results in a decrease in engraftment, an increase in long-term HSC frequency and loss of stem cell polarity. Exposure of aged HSCs to thrombin-cleaved OPN attenuates aging of old HSCs, resulting in increased engraftment, decreased HSC frequency, increased stem cell polarity and a restored balance of lymphoid and myeloid cells in peripheral blood. Thus, our data suggest a critical role for reduced stroma-derived OPN for HSC aging and identify thrombin-cleaved OPN as a novel niche informed therapeutic approach for ameliorating HSC phenotypes associated with aging.
publishedVersion

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::36c6b80412477038cc8e3c62bc7c543bTest
http://europepmc.org/articles/PMC5376966Test

المؤلفون: Vijay P.S. Rawat, Stefanie Sieste, Anne Stumper, Naidu M. Vegi, Christian Buske, Tanja Weil, Michaela Feuring-Buske, Maksymilian Marek Zegota, Seah Ling Kuan, Sabyasachi Chakrabortty, Sven Rau

المصدر: Scientific Reports
Scientific Reports, Vol 10, Iss 1, Pp 1-10 (2020)

مصطلحات موضوعية: 0301 basic medicine, Adult, Male, Myeloid, HL60, CD34, lcsh:Medicine, Apoptosis, Ruthenium, Article, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Drug Stability, hemic and lymphatic diseases, Cell Line, Tumor, medicine, Humans, Leukaemia, Receptors, Somatostatin, Clonogenic assay, lcsh:Science, Aged, Multidisciplinary, Photosensitizing Agents, Somatostatin receptor, Chemistry, lcsh:R, Myeloid leukemia, Middle Aged, medicine.disease, Fetal Blood, Leukemia, Leukemia, Myeloid, Acute, 030104 developmental biology, Somatostatin, medicine.anatomical_structure, Photochemotherapy, 030220 oncology & carcinogenesis, Drug delivery, Cancer research, lcsh:Q, Female, Lysosomes, Reactive Oxygen Species

الوصف: Acute myeloid leukemia (AML) is characterized by relapse and treatment resistance in a major fraction of patients, underlining the need of innovative AML targeting therapies. Here we analysed the therapeutic potential of an innovative biohybrid consisting of the tumor-associated peptide somatostatin and the photosensitizer ruthenium in AML cell lines and primary AML patient samples. Selective toxicity was analyzed by using CD34 enriched cord blood cells as control. Treatment of OCI AML3, HL60 and THP1 resulted in a 92, and 99 and 97% decrease in clonogenic growth compared to the controls. Primary AML cells demonstrated a major response with a 74 to 99% reduction in clonogenicity in 5 of 6 patient samples. In contrast, treatment of CD34+ CB cells resulted in substantially less reduction in colony numbers. Subcellular localization assays of RU-SST in OCI-AML3 cells confirmed strong co-localization of RU-SST in the lysosomes compared to the other cellular organelles. Our data demonstrate that conjugation of a Ruthenium complex with somatostatin is efficiently eradicating LSC candidates of patients with AML. This indicates that receptor mediated lysosomal accumulation of photodynamic metal complexes is a highly attractive approach for targeting AML cells.

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الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::87a7489c054182da6f01e5caefc9e2f6Test
https://pubmed.ncbi.nlm.nih.gov/31941913Test

المؤلفون: Theo Lasser, Lorna Hodgson, Paul Verkade, Marco Raabe, Tanja Weil, Katharina Landfester, Judith Mantell, Ingo Lieberwirth, Shen Han

المصدر: Nano Letters
Han, S, Raabe, M, Hodgson, L, Mantell, J, Verkade, P, Lasser, T, Landfester, K, Weil, T & Lieberwirth, I 2019, ' High-Contrast Imaging of Nanodiamonds in Cells by Energy Filtered and Correlative Light-Electron Microscopy : Toward a Quantitative Nanoparticle-Cell Analysis ', Nano Letters, vol. 19, no. 3, pp. 2178-2185 . https://doi.org/10.1021/acs.nanolett.9b00752Test

مصطلحات موضوعية: Letter, particle-cell interactions, Nanoparticle, Contrast Media, nanodiamond, Cell analysis, 02 engineering and technology, localization, law.invention, law, energy filtered transmission electron microscopy, Microscopy, General Materials Science, Nanodiamond, Instrumentation, 0303 health sciences, Vesicle, High contrast imaging, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Fluorescence, nanoparticle quantification, Cell Tracking, Drug delivery, delivery, 0210 nano-technology, Correlative, Materials science, Bioengineering, Nanotechnology, Electrons, Nanodiamonds, 03 medical and health sciences, protein corona, Energy filtered transmission electron microscopy, Humans, fluorescent nanodiamonds, 030304 developmental biology, Fluorescent Dyes, Organelles, Mechanical Engineering, Correlative light-electron microscopy, General Chemistry, energy-filtered transmission electron microscopy, Nanostructures, Microscopy, Electron, Biophysics, Correlative light electron microscopy, Electron microscope, probes, Energy (signal processing), HeLa Cells

الوصف: Fluorescent nanodiamonds (fNDs) represent an emerging class of nanomaterials offering great opportunities for ultrahigh resolution imaging, sensing and drug delivery applications. Their biocompatibility, exceptional chemical and consistent photostability renders them particularly attractive for correlative light-electron microscopy studies providing unique insights into nanoparticle-cell interactions. Herein, we demonstrate a stringent procedure to image and quantify fNDs with a high contrast down to the single particle level in cells. Individual fNDs were directly visualized by energy-filtered transmission electron microscopy, that is, inside newly forming, early endosomal vesicles during their cellular uptake processes as well as inside cellular organelles such as a mitochondrion. Furthermore, we demonstrate the unequivocal identification, localization, and quantification of individual fNDs in larger fND clusters inside intracellular vesicles. Our studies are of great relevance to obtain quantitative information on nanoparticle trafficking and their various interactions with cells, membranes, and organelles, which will be crucial to design-improved sensors, imaging probes, and nanotherapeutics based on quantitative data.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::53eea8905b41e183fdd035b660d37146Test
https://pubmed.ncbi.nlm.nih.gov/30810045Test

المؤلفون: Thomas Mack, Stefanie Sieste, Bernd Knöll, Francesco Simone Ruggeri, Ramin Naraghi, Christopher V. Synatschke, Tristan Bereau, Tuomas P. J. Knowles, Corinna Schilling, Tomas Sneideris, Tanja Weil, Selene Lickfett, Arghya Dutta, Daniela Sinske

المصدر: Advanced Functional Materials, 29(24)
Advanced Functional Materials 29 (2019) 24
Advanced Functional Materials

مصطلحات موضوعية: Materials science, Peptide, 02 engineering and technology, Biomaterials, 03 medical and health sciences, Electrochemistry, 030304 developmental biology, Sequence (medicine), chemistry.chemical_classification, 0303 health sciences, self-assembling peptide, Regeneration (biology), neuron growth, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Electronic, Optical and Magnetic Materials, Peripheral, Cell biology, chemistry, Nanofiber, regeneration, facial nerve, 0210 nano-technology, naofiber, Self-assembling peptide

الوصف: There is an urgent need for biomaterials that support tissue healing, particularly neuronal regeneration. In a medium throughput screen novel self-assembling peptide (SAP) sequences that form fibrils and stimulated nerve fiber growth of peripheral nervous system (PNS)-derived neurons are identified. Based on the peptide sequences and fibril morphologies and by applying rational data-mining, important structural parameters stimulating neuronal activity are elucidated. Three SAPs (SAP1e, SAP2e, and SAP5c) enhance adhesion and growth of PNS neurons. These SAPs form 2D and 3D matrices that serve as bioactive scaffolds stimulating cell adhesion and growth. The newly discovered SAPs also support the growth of CNS neurons and glia cells. Subsequently, the potential of SAPs to enhance PNS regeneration in vivo is analyzed. For this, the facial nerve driving whisker movement in mice is injured. Notably, SAPs persist for up to 3 weeks in the injury site indicating highly adhesive properties and stability. After SAP administration, more motor neurons incorporating markers for successive regeneration are observed. Recovery of whisker movement is elevated in SAP-injected mice. In summary, short peptides that form fibrils are identified and the adhesion, growth, and regeneration of neurons have been efficiently enhanced without the necessity to attach hormones or growth factors.

وصف الملف: application/octet-stream; text/html; application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4e215ecbe8e3d761041f6332e137ea25Test
https://research.wur.nl/en/publications/sequence-optimized-peptide-nanofibers-as-growth-stimulators-for-rTest

المؤلفون: David Y. W. Ng, Seah Ling Kuan, Tanja Weil, Stephan Fischer, Kaloian Koynov, Weina Liu, Yu Tokura, Christina Förtsch, Yuzhou Wu, Holger Barth

المصدر: Macromolecular Bioscience. 16:803-810

مصطلحات موضوعية: 0301 basic medicine, Streptavidin, Dendrimers, Botulinum Toxins, Polymers and Plastics, Supramolecular chemistry, Biotin, Bioengineering, 010402 general chemistry, medicine.disease_cause, 01 natural sciences, Cell Line, law.invention, Biomaterials, 03 medical and health sciences, chemistry.chemical_compound, Cytosol, Confocal microscopy, law, Clostridium botulinum, Materials Chemistry, medicine, Humans, Serum Albumin, Toxins, Biological, ADP Ribose Transferases, chemistry.chemical_classification, Chemistry, Macrophages, Transporter, Human serum albumin, 0104 chemical sciences, 030104 developmental biology, Enzyme, Biochemistry, Biotechnology, medicine.drug

الوصف: Nature has provided a highly optimized toolbox in bacterial endotoxins with precise functions dictated by their clear structural division. Inspired by this streamlined design, a supramolecular approach capitalizing on the strong biomolecular (streptavidin (SA))-biotin interactions is reported herein to prepare two multipartite fusion constructs, which involves the generation 2.0 (D2) or generation 3.0 (D3) polyamidoamine-dendronized transporter proteins (dendronized streptavidin (D3SA) and dendronized human serum albumin (D2HSA)) non-covalently fused to the C3bot1 enzyme from Clostridium botulinum, a potent and specific Rho-inhibitor. The fusion constructs, D3SA-C3 and D2HSA-C3, represent the first examples of dendronized protein transporters that are fused to the C3 enzyme, and it is successfully demonstrated that the C3 Rho-inhibitor is delivered into the cytosol of mammalian cells as determined from the characteristic C3-mediated changes in cell morphology and confocal microscopy. The design circumvents the low uptake of the C3 enzyme by eukaryotic cells and holds great promise for reprogramming the properties of toxin enzymes using a supramolecular approach to broaden their therapeutic applications.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::572e6c09e94f5279751084a70ee0f212Test
https://doi.org/10.1002/mabi.201500417Test

المؤلفون: Malin Insa Jansen, Jana Hedrich, Heiko J. Luhmann, Tanja Weil, David Y. W. Ng, Pierpaolo Moscariello

المصدر: Advanced Science

مصطلحات موضوعية: 0301 basic medicine, Streptavidin, blood‐brain barrier, General Chemical Engineering, General Physics and Astronomy, Medicine (miscellaneous), 02 engineering and technology, Blood–brain barrier, Biochemistry, Genetics and Molecular Biology (miscellaneous), 03 medical and health sciences, chemistry.chemical_compound, In vivo, Dendrimer, medicine, streptavidin, General Materials Science, PAMAM dendrimer, Full Paper, General Engineering, Full Papers, 021001 nanoscience & nanotechnology, In vitro, 3. Good health, 030104 developmental biology, medicine.anatomical_structure, Transcytosis, chemistry, Biotinylation, Drug delivery, drug delivery, Biophysics, 0210 nano-technology

الوصف: Neurological disorders are undoubtedly among the most alarming diseases humans might face. In treatment of neurological disorders, the blood‐brain barrier (BBB) is a challenging obstacle preventing drug penetration into the brain. Advances in dendrimer chemistry for central nervous system (CNS) treatments are presented here. A poly(amido)amine (PAMAM) dendrimer bioconjugate with a streptavidin adapter for the attachment of dendrons or any biotinylated drug is constructed. In vitro studies on porcine or murine models and in vivo mouse studies are performed and reveal the permeation of dendronized streptavidin (DSA) into the CNS. The bioconjugate is taken up mainly by the caveolae pathway and transported across the BBB via transcytosis escaping from lysosomes. After transcytosis DSA are delivered to astrocytes and neurons. Furthermore, DSA offer high biocompatibility in vitro and in vivo. In summary, a new strategy for implementing therapeutic PAMAM function as well as drug delivery in neuropathology is presented here.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::db501171c8d9728a21fff7f506a766b0Test
http://europepmc.org/articles/PMC5979778Test