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Institute of Solid-State Physics, University of Latvia, Latvia
Cellbox labs LTD, Atari
Faculty of Materials Science and Applied Chemistry, Institute of General Chemical Engineering, Riga Technical university, Latvia
Export Date: 5 March 2024
Correspondence Address: Bajo-Santos, C.; Latvian Biomedical Research and Study CentreLatvia; email: cristina.bajo@biomed.lu.lv
Chemicals/CAS: Polymers
Funding details: 739508
Funding details: Latvijas Zinātnes Padome, lzp-2019/1-0142, lzp-2022/1-0373
Funding text 1: We thank all the donors who participated in this study, the staff of the Latvian Genome Database for providing the samples. The Institute of Solid-State Physics, University of Latvia as the Center of Excellence has received funding from the European Union's Horizon 2020 Framework Programme H2020-WIDESPREAD-01-2016-2017-TeamongPhase2 under grant agreement No. 739508, project CAMART2. This work was supported by The Latvian Council of Science Project No. lzp-2019/1-0142 and Project No: lzp-2022/1-0373.
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Extracellular vesicles (EVs) hold immense potential for various biomedical applications, including diagnostics, drug delivery, and regenerative medicine. Nevertheless, the current methodologies for isolating EVs present significant challenges, such as complexity, time consumption, and the need for bulky equipment, which hinders their clinical translation. To address these limitations, we aimed to develop an innovative microfluidic system based on cyclic olefin copolymer-off-stoichiometry thiol-ene (COC-OSTE) for the efficient isolation of EVs from large-volume samples in a continuous manner. By utilizing size and buoyancy-based separation, the technology used in this study achieved a significantly narrower size distribution compared to existing approaches from urine and cell media samples, enabling the targeting of specific EV size fractions in future applications. Our innovative COC-OSTE microfluidic device design, utilizing bifurcated asymmetric flow field-flow fractionation technology, offers a straightforward and continuous EV isolation approach for large-volume samples. Furthermore, the potential for mass manufacturing of this microfluidic device offers scalability and consistency, making it feasible to integrate EV isolation into routine clinical diagnostics and industrial processes, where high consistency and throughput are essential requirements. © 2024 JoVE Journal of Visualized Experiments.
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<pre class="comment" style="margin-top: 0; margin-bottom: 0;"><u>Megjegyzés</u>: Latvian Biomedical Research and Study Centre, Latvia
Institute of Solid-State Physics, University of Latvia, Latvia
Cellbox labs LTD, Atari
Faculty of Materials Science and Applied Chemistry, Institute of General Chemical Engineering, Riga Technical university, Latvia
Export Date: 5 March 2024 ...</pre>
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Correspondence Address: Eduati, F.; Department of Biomedical Engineering, Netherlands; email: f.eduati@tue.nl
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Funding text 1: We would like to thank Stacey Martina of the NanoLab TuE for help with HMDS vapour deposition. This research was funded by the Institute for Complex Molecular Systems (ICMS) at TU/ e and by the Netherlands Organization for Scientific Research (NWO) Gravitation programme IMAGINE! (project number 24.005.009).
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Droplet microfluidics is a versatile tool that allows the execution of a large number of reactions in chemically distinct nanoliter compartments. Such systems have been used to encapsulate a variety of biochemical reactions-from incubation of single cells to implementation of PCR reactions, from genomics to chemical synthesis. Coupling the microfluidic channels with regulatory valves allows control over their opening and closing, thereby enabling the rapid production of large-scale combinatorial libraries consisting of a population of droplets with unique compositions. In this paper, protocols for the fabrication and operation of a pressure-driven, PDMS-based bilayer microfluidic device that can be utilized to generate combinatorial libraries of water-in-oil emulsions called plugs are presented. By incorporating software programs and microfluidic hardware, the flow of desired fluids in the device can be controlled and manipulated to generate combinatorial plug libraries and to control the composition and quantity of constituent plug populations. These protocols will expedite the process of generating combinatorial screens, particularly to study drug response in cells from cancer patient biopsies. © 2023 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.
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<pre class="comment" style="margin-top: 0; margin-bottom: 0;"><u>Megjegyzés</u>: Export Date: 19 February 2024
Correspondence Address: Eduati, F.; Department of Biomedical Engineering, Netherlands; email: f.eduati@tue.nl
Funding details: Nederlandse Organisatie voor Wetenschappelijk Onderzoek, NWO, 24.005.009
Funding text 1: We would like to thank Stacey Martina of the NanoLab TuE for help with HMDS vap...</pre>
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Most bacteria, including mycobacteria, generate extracellular vesicles (EVs). Since bacterial EVs (bEVs) contain a subset of cellular components, including metabolites, lipids, proteins, and nucleic acids, several groups have evaluated either the native or recombinant versions of bEVs for their protective potency as subunit vaccine candidates. Unlike native EVs, recombinant EVs are molecularly engineered to contain one or more immunogens of interest. Over the last decade, different groups have explored diverse approaches for generating recombinant bEVs. However, here, we report the design, construction, and enrichment of recombinant mycobacterial EVs (mEVs) in mycobacteria. Towards that, we use Mycobacterium smegmatis (Msm), an avirulent soil mycobacterium as the model system. We first describe the generation and enrichment of native EVs of Msm. Then, we describe the design and construction of recombinant mEVs that contain either mCherry, a red fluorescent reporter protein, or EsxA (Esat-6), a prominent immunogen of Mycobacterium tuberculosis. We achieve this by separately fusing mCherry and EsxA N-termini with the C-terminus of a small Msm protein Cfp-29. Cfp-29 is one of the few abundantly present proteins of MsmEVs. The protocol to generate and enrich recombinant mEVs from Msm remains identical to the generation and enrichment of native EVs of Msm.
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<div class="JournalArticle Publication short-list"> <div class="authors"> <span class="author-name" > Jayaswal, Praapti </span> <span class="author-type"> </span> ; <span class="author-name" > Ilyas, Mohd </span> <span class="author-type"> </span> ; <span class="author-name" > Singh, Kuljit </span> <span class="author-type"> </span> ; <span class="author-name" > Kumar, Saurabh </span> <span class="author-type"> </span> ; <span class="author-name" > Sisodiya, Lovely </span> <span class="author-type"> </span> ; <span class="author-name" > Jain, Sapna </span> <span class="author-type"> </span> ; <span class="author-name" > Mahlawat, Rahul </span> <span class="author-type"> </span> ; <span class="author-name" > Sharma, Nishant </span> <span class="author-type"> </span> ; <span class="author-name" > Gupta, Vishal </span> <span class="author-type"> </span> ; <span class="author-name" > Atmakuri, Krishnamohan ✉ </span> <span class="author-type"> </span> </div ><div class="title"><a href="/gui2/?mode=browse¶ms=publication;34613123" mtid="34613123" target="_blank">Enrichment of Native and Recombinant Extracellular Vesicles of Mycobacteria</a></div> <div class="pub-info"> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span> <span class="journal-volume"></span> : <span class="journal-issue">202</span> <span class="page"> Paper: e65138 , 22 p. </span> <span class="year">(2023)</span> </div> <div class="pub-end"><div class="identifier-list"> <span class="identifiers"> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="10.3791/65138" target="_blank" href="https://doi.org/10.3791/65138"> DOI </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="001127859900002" target="_blank" href="https://www.webofscience.com/wos/woscc/full-record/001127859900002"> WoS </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="85179314577" target="_blank" href="http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-85179314577"> Scopus </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="38145372" target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=38145372&dopt=Abstract"> PubMed </a> </span> </span> </div> <div class="short-pub-prop-list"> <span class="short-pub-mtid"> Közlemény:34613123 </span> <span class="status-holder"><span class="status-data status-VALIDATED"> Egyeztetett </span></span> <span class="pub-core"> Idéző </span> <span class="pub-type">Folyóiratcikk (Szakcikk ) </span> <!-- && !record.category.scientific --> <span class="pub-category">Tudományos</span> </div> </div> </div><div class="JournalArticle Publication long-list">
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Vibrational sum-frequency generation (VSFG), a second-order nonlinear optical signal, has traditionally been used to study molecules at interfaces as a spectroscopy technique with a spatial resolution of -100 mu m. However, the spectroscopy is not sensitive to the heterogeneity of a sample. To study mesoscopically heterogeneous samples, we, along with others, pushed the resolution limit of VSFG spectroscopy down to -1 mu m level and constructed the VSFG microscope. This imaging technique not only can resolve sample morphologies through imaging, but also record a broadband VSFG spectrum at every pixel of the images. Being a second-order nonlinear optical technique, its selection rule enables the visualization of noncentrosymmetric or chiral self-assembled structures commonly found in biology, materials science, and bioengineering, among others. In this article, the audience will be guided through an inverted transmission design that allows for imaging unfixed samples. This work also showcases that VSFG microscopy can resolve chemicalspecific geometric information of individual self-assembled sheets by combining it with a neural network function solver. Lastly, the images obtained under brightfield, SHG, and VSFG configurations of various samples briefly discuss the unique information revealed by VSFG imaging.
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<div class="JournalArticle Publication short-list"> <div class="authors"> <span class="author-name" > Wagner, Jackson C. </span> <span class="author-type"> </span> ; <span class="author-name" > Yang, Bin </span> <span class="author-type"> </span> ; <span class="author-name" > Wu, Zishan </span> <span class="author-type"> </span> ; <span class="author-name" > Xiong, Wei ✉ </span> <span class="author-type"> </span> </div ><div class="title"><a href="/gui2/?mode=browse¶ms=publication;34611670" mtid="34611670" target="_blank">Multimodal Nonlinear Hyperspectral Chemical Imaging Using Line-Scanning Vibrational Sum-Frequency Generation Microscopy</a></div> <div class="pub-info"> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span> <span class="journal-volume"></span> : <span class="journal-issue">202</span> <span class="page"> Paper: e65388 , 18 p. </span> <span class="year">(2023)</span> </div> <div class="pub-end"><div class="identifier-list"> <span class="identifiers"> <span class="id identifier oa_none" title="none"> <a style="color:black" title="10.3791/65388" target="_blank" href="https://doi.org/10.3791/65388"> DOI </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="001127859900021" target="_blank" href="https://www.webofscience.com/wos/woscc/full-record/001127859900021"> WoS </a> </span> </span> </div> <div class="short-pub-prop-list"> <span class="short-pub-mtid"> Közlemény:34611670 </span> <span class="status-holder"><span class="status-data status-VALIDATED"> Egyeztetett </span></span> <span class="pub-core"> Idéző </span> <span class="pub-type">Folyóiratcikk (Szakcikk ) </span> <!-- && !record.category.scientific --> <span class="pub-category">Tudományos</span> </div> </div> </div><div class="JournalArticle Publication long-list">
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Correspondence Address: Didonna, A.; Department of Anatomy and Cell Biology, United States; email: didonnaal21@ecu.edu
Chemicals/CAS: Autoantibodies; Cytokines
Funding details: National Institutes of Health, NIH, R03NS131908
Funding details: U.S. Department of Defense, DOD, W81XWH-22-1-0517
Funding details: East Carolina University, ECU
Funding text 1: This study was supported by the National Institutes of Health (R03NS131908) and the Department of Defense through the Multiple Sclerosis Research Program under Award No. W81XWH-22-1-0517. Opinions, interpretations, conclusions and recommendations are those of the author and are not necessarily endorsed by the Department of Defense. This study was also supported by East Carolina University startup funds.
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Experimental autoimmune encephalomyelitis (EAE) is a disease model that recapitulates the autoimmune disorder multiple sclerosis (MS) at histopathological and molecular levels. EAE is induced by immunizing experimental animals via subcutaneous injection of short myelin peptides together with specific adjuvants to boost the immune response. Like the human counterpart, EAE mice develop demyelinating lesions, immune cell infiltration into the central nervous system (CNS), glia activation and neuronal injury. A consistent body of evidence also supports a mechanistic role for B cell dysfunction in the etiology of both MS and EAE. B cells can serve as antigen-presenting cells as well as a primary source of pro-inflammatory cytokines and autoantibodies. In EAE, antibodies are generated against the myelin peptides that were employed to induce the disease. Such autoantibodies have been shown to mediate either myelin loss or pathogenic T cell reactivation into the CNS. This article describes an efficient ELISA-based protocol to quantify autoantibodies in the serum of C57BL/6J mice immunized with the myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) peptide. The proposed method serves as a powerful tool to investigate the specificity and magnitude of the aberrant humoral response in the context of autoimmune demyelination. © 2023 JoVE Journal of Visualized Experiments.
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<div class="JournalArticle Publication short-list"> <div class="authors"> <span class="author-name" > Carver, J.J. </span> <span class="author-type"> </span> ; <span class="author-name" > Didonna, A. ✉ </span> <span class="author-type"> </span> </div ><div class="title"><a href="/gui2/?mode=browse¶ms=publication;34564284" mtid="34564284" target="_blank">Quantification of Autoreactive Antibodies in Mice upon Experimental Autoimmune Encephalomyelitis</a></div> <div class="pub-info"> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span> <span class="journal-volume">2023</span> : <span class="journal-issue">202</span> <span class="page"> Paper: 66218 </span> <span class="year">(2023)</span> </div> <div class="pub-end"><div class="identifier-list"> <span class="identifiers"> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="10.3791/66218" target="_blank" href="https://doi.org/10.3791/66218"> DOI </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="001127859900010" target="_blank" href="https://www.webofscience.com/wos/woscc/full-record/001127859900010"> WoS </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:black" title="85179970611" target="_blank" href="http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-85179970611"> Scopus </a> </span> </span> </div> <div class="short-pub-prop-list"> <span class="short-pub-mtid"> Közlemény:34564284 </span> <span class="status-holder"><span class="status-data status-VALIDATED"> Egyeztetett </span></span> <span class="pub-core"> Idéző </span> <span class="pub-type">Folyóiratcikk (Szakcikk ) </span> <!-- && !record.category.scientific --> <span class="pub-category">Tudományos</span> </div> </div> </div><div class="JournalArticle Publication long-list">
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<div class="title"><a href="/gui2/?mode=browse¶ms=publication;34564284" target="_blank">Quantification of Autoreactive Antibodies in Mice upon Experimental Autoimmune Encephalomyelitis</a></div> <div> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span>
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<div class="lastModified">Utolsó módosítás: 2024.02.07. 12:40 Tóth Andrea (SZTE admin5 ÁOK Neurológiai Klinika)
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<pre class="comment" style="margin-top: 0; margin-bottom: 0;"><u>Megjegyzés</u>: Export Date: 7 February 2024
Correspondence Address: Didonna, A.; Department of Anatomy and Cell Biology, United States; email: didonnaal21@ecu.edu
Chemicals/CAS: Autoantibodies; Cytokines
Funding details: National Institutes of Health, NIH, R03NS131908
Funding details: U.S. Department of Defense, DO...</pre>
</div></div>
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Key Laboratory of Medical Electrophysiology of Ministry of Education, Collaborative Innovation Center for Prevention and Treatment of Cardiovascular Disease, Institute of Cardiovascular Research, Southwest Medical University, China
Department of Cardiology, the Affiliated Hospital of Southwest Medical University, China
Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, China
Department of Pharmacology, University of Oxford, United Kingdom
Export Date: 31 January 2024
Correspondence Address: Tan, X.; Key Laboratory of Medical Electrophysiology of Ministry of Education, China; email: tanxiaoqiu@swmu.edu.cn
Correspondence Address: Lei, M.; Key Laboratory of Medical Electrophysiology of Ministry of Education, China; email: ming.lei@pharm.ox.ac.uk
Correspondence Address: Ou, X.; Key Laboratory of Medical Electrophysiology of Ministry of Education, China; email: oxh8081@swmu.edu.cn
Chemicals/CAS: calcium, 7440-70-2, 14092-94-5; Calcium; Ryanodine Receptor Calcium Release Channel
Funding details: Sichuan Province Science and Technology Support Program, 2021YJ0206, 2022NSFSC1602, 2022YFS0607, 23ZYZYTS0433
Funding details: Medical Research Council, MRC, G1002082, G10031871181, ML02647
Funding details: British Heart Foundation, BHF, PG/11/59/29004 ML, PG/12/21/29473, PG/14/80/31106, PG/16/67/32340
Funding details: National Natural Science Foundation of China, NSFC, 31871181, 81700308, 82270334
Funding details: Guangxi Normal University, GXNU, CMEMR2017-B08
Funding details: Key Laboratory of Bioorganic Chemistry and Molecular Engineering, Ministry of Education
Funding text 1: This study is supported by the National Natural Science Foundation of China (81700308 to XO and 31871181 to ML, and 82270334 to XT), Sichuan Province Science and Technology Support Program (CN) (2021YJ0206 to XO, 23ZYZYTS0433, and 2022YFS0607 to XT, and 2022NSFSC1602 to TC) and State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources (Guangxi Normal University) (CMEMR2017-B08 to XO), MRC (G10031871181 to ML02647, G1002082, ML), BHF (PG/14/80/31106, PG/16/67/32340, PG/12/21/29473, PG/11/59/29004 ML), BHF CRE at Oxford (ML) grants.
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Li, Y.
Dual-Dye Optical Mapping of Hearts from RyR2R2474S Knock-In Mice of Catecholaminergic Polymorphic Ventricular Tachycardia
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The pro-arrhythmic cardiac disorder catecholaminergic polymorphic ventricular tachycardia (CPVT) manifests as polymorphic ventricular tachycardia episodes following physical activity, stress, or catecholamine challenge, which can deteriorate into potentially fatal ventricular fibrillation. The mouse heart is a widespread species for modeling inherited cardiac arrhythmic diseases, including CPVT. Simultaneous optical mapping of transmembrane potential (Vm) and calcium transients (CaT) from Langendorff-perfused mouse hearts has the potential to elucidate mechanisms underlying arrhythmogenesis. Compared with the cellular level investigation, the optical mapping technique can test some electrophysiological parameters, such as the determination of activation, conduction velocity, action potential duration, and CaT duration. This paper presents the instrumentation setup and experimental procedure for high-throughput optical mapping of CaT and Vm in murine wild-type and heterozygous RyR2-R2474S/+ hearts, combined with programmed electrical pacing before and during the isoproterenol challenge. This approach has demonstrated a feasible and reliable method for mechanistically studying CPVT disease in an ex vivo mouse heart preparation. © 2023 JoVE Journal of Visualized Experiments.
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<div class="JournalArticle Publication short-list"> <div class="authors"> <span class="author-name" > Li, Y. </span> <span class="author-type"> </span> ; <span class="author-name" > Yang, J. </span> <span class="author-type"> </span> ; <span class="author-name" > Zhang, R. </span> <span class="author-type"> </span> ; <span class="author-name" > Chen, T. </span> <span class="author-type"> </span> ; <span class="author-name" > Zhang, S. </span> <span class="author-type"> </span> ; <span class="author-name" > Zheng, Y. </span> <span class="author-type"> </span> ; <span class="author-name" > Wen, Q. </span> <span class="author-type"> </span> ; <span class="author-name" > Li, T. </span> <span class="author-type"> </span> ; <span class="author-name" > Tan, X. ✉ </span> <span class="author-type"> </span> ; <span class="author-name" > Lei, M. ✉ </span> <span class="author-type"> </span> et al. </div ><div class="title"><a href="/gui2/?mode=browse¶ms=publication;34547956" mtid="34547956" target="_blank">Dual-Dye Optical Mapping of Hearts from RyR2R2474S Knock-In Mice of Catecholaminergic Polymorphic Ventricular Tachycardia</a></div> <div class="pub-info"> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span> <span class="journal-volume">2023</span> : <span class="journal-issue">202</span> <span class="page"> Paper: 65082 </span> <span class="year">(2023)</span> </div> <div class="pub-end"><div class="identifier-list"> <span class="identifiers"> <span class="id identifier oa_none" title="none"> <a style="color:black" title="10.3791/65082" target="_blank" href="https://doi.org/10.3791/65082"> DOI </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:black" title="85180674644" target="_blank" href="http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-85180674644"> Scopus </a> </span> </span> </div> <div class="short-pub-prop-list"> <span class="short-pub-mtid"> Közlemény:34547956 </span> <span class="status-holder"><span class="status-data status-APPROVED"> Nyilvános </span></span> <span class="pub-core"> Idéző </span> <span class="pub-type">Folyóiratcikk (Szakcikk ) </span> <!-- && !record.category.scientific --> <span class="pub-category">Tudományos</span> </div> </div> </div><div class="JournalArticle Publication long-list">
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<div class="title"><a href="/gui2/?mode=browse¶ms=publication;34547956" target="_blank">Dual-Dye Optical Mapping of Hearts from RyR2R2474S Knock-In Mice of Catecholaminergic Polymorphic Ventricular Tachycardia</a></div> <div> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span>
<span class="journal-issn">(<a target="_blank" href="https://portal.issn.org/resource/ISSN/1940-087X">1940-087X</a> <a target="_blank" href="https://portal.issn.org/resource/ISSN/1940-087X">1940-087X</a>)</span>:
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<div class="lastModified">Utolsó módosítás: 2024.01.31. 11:37 Sonnevend Kinga (SE_AOK_Adm5_Élet_kutcsop, admin)
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<pre class="comment" style="margin-top: 0; margin-bottom: 0;"><u>Megjegyzés</u>: Key Laboratory of Medical Electrophysiology of Ministry of Education, Collaborative Innovation Center for Prevention and Treatment of Cardiovascular Disease, Institute of Cardiovascular Research, Southwest Medical University, China
Department of Cardiology, the Affiliated Hospital of Southwest Medical University, China
Department of Cardiology, Un...</pre>
</div></div>
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Sample preparation is crucial for elemental determination, and various techniques are available, one of which involves homogenization followed by acid digestion. Special care is required during sample handling in the preparation stage to eliminate or minimize potential contamination and analyte loss. Homogenization is a process that simultaneously reduces particle size and uniformly distributes sample components. Following homogenization, the sample undergoes acid digestion, wherein it is digested with acids and auxiliary chemicals at elevated temperatures, transforming solid samples into a liquid state. In this process, metals in the original sample react with acids to form water-soluble salts. Samples prepared through acid digestion are suitable for elemental analysis using techniques such as inductively coupled plasma mass spectrometry, inductively coupled plasma optical emission spectroscopy, atomic absorption spectroscopy, electrochemical methods, and other analytical techniques. This work details the preparation of food samples for multi-element determination using inductively coupled plasma mass spectrometry. The step-by-step procedure involves the homogenization process using a laboratory-sized mixer with ceramic blades, followed by acid digestion in closed vessels using microwave-assisted wet acid digestion. A mixture of 5.0 mL of 68 wt% HNO3 and 1.0 mL of 30 wt% H2O2 serves as an auxiliary reagent. This guide provides an explanation of the processes involved in both stages.
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<div class="JournalArticle Publication short-list"> <div class="authors"> <span class="author-name" > Rantaša, Matjaž </span> <span class="author-type"> </span> ; <span class="author-name" > Majer, David </span> <span class="author-type"> </span> ; <span class="author-name" > Finšgar, Matjaž </span> <span class="author-type"> </span> </div ><div class="title"><a href="/gui2/?mode=browse¶ms=publication;34547801" mtid="34547801" target="_blank">Preparation of Food Samples Using Homogenization and Microwave-Assisted Wet Acid Digestion for Multi-Element Determination with ICP-MS</a></div> <div class="pub-info"> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span> <span class="journal-volume">2023</span> : <span class="journal-issue">202</span> <span class="page"> Paper: e65624 </span> <span class="year">(2023)</span> </div> <div class="pub-end"><div class="identifier-list"> <span class="identifiers"> <span class="id identifier oa_none" title="none"> <a style="color:black" title="10.3791/65624" target="_blank" href="https://doi.org/10.3791/65624"> DOI </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="85180733545" target="_blank" href="http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-85180733545"> Scopus </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="38189412" target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=38189412&dopt=Abstract"> PubMed </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:black" title="https://www.jove.com/t/65624/preparation-food-samples-using-homogenization-microwave-assisted-wet" target="_blank" href="https://www.jove.com/t/65624/preparation-food-samples-using-homogenization-microwave-assisted-wet"> Egyéb URL </a> </span> </span> </div> <div class="short-pub-prop-list"> <span class="short-pub-mtid"> Közlemény:34547801 </span> <span class="status-holder"><span class="status-data status-APPROVED"> Nyilvános </span></span> <span class="pub-core"> Idéző </span> <span class="pub-type">Folyóiratcikk (Szakcikk ) </span> <!-- && !record.category.scientific --> <span class="pub-category">Tudományos</span> </div> </div> </div><div class="JournalArticle Publication long-list"> <div class="authors"> <img title="Idézőközlemény" style="float: left" src="/frontend/resources/grid/publication-citation-icon.png"> <div class="autype autype0"> <span class="author-name" >Rantaša Matjaž </span> ; 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Department of Biochemistry and Molecular Pharmacology, NYU Grossman School of Medicine, United States
The Laura and Isaac Perlmutter Cancer Center, NYU Langone Health, United States
Department of Cell Biology, NYU Grossman School of Medicine, United States
Howard Hughes Medical Institute, NYU Grossman School of Medicine, United States
Institute of Enzymology, Centre of Excellence of the Hungarian Academy of Sciences, HUN-REN Research Centre for Natural Sciences, United States
Export Date: 9 January 2024
Correspondence Address: Rona, G.; Department of Biochemistry and Molecular Pharmacology, United States; email: gergely.rona@nyulangone.org
Funding details: National Institutes of Health, NIH, GM136250
Funding text 1: The authors thank Michele Pagano for his support and helpful insights, Ashley Chui and Sharon Kaisari for critically reading the manuscript, and Jeffrey Estrada and Vilma Diaz for their continuous support. This work was supported by a diversity supplement to the National Institutes of Health grant GM136250.
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DNA has dedicated cellular repair pathways capable of coping with lesions that could arise from both endogenous and/or exogenous sources. DNA repair necessitates collaboration between numerous proteins, responsible for covering a broad range of tasks from recognizing and signaling the presence of a DNA lesion to physically repairing it. During this process, tracks of single-stranded DNA (ssDNA) are often created, which are eventually filled by DNA polymerases. The nature of these ssDNA tracks (in terms of both length and number), along with the polymerase recruited to fill these gaps, are repair pathway-specific. The visualization of these ssDNA tracks can help us understand the complicated dynamics of DNA repair mechanisms. This protocol provides a detailed method for the preparation of G1 synchronized cells to measure ssDNA foci formation upon genotoxic stress. Using an easy-toutilize immunofluorescence approach, we visualize ssDNA by staining for RPA2, a component of the heterotrimeric replication protein A complex (RPA). RPA2 binds to and stabilizes ssDNA intermediates that arise upon genotoxic stress or replication to control DNA repair and DNA damage checkpoint activation. 5-Ethynyl-2'-deoxyuridine (EdU) staining is used to visualize DNA replication to exclude any S phase cells. This protocol provides an alternative approach to the conventional, non-denaturing 5-bromo-2'-deoxyuridine (BrdU)-based assays and is better suited for the detection of ssDNA foci outside the S phase. © 2023 JoVE Creative Commons Attribution.
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<div class="JournalArticle Publication short-list"> <div class="authors"> <span class="author-name" > Zhang, Q. </span> <span class="author-type"> </span> ; <span class="author-name" > Kerzhnerman, M.A. </span> <span class="author-type"> </span> ; <span class="author-name" > García-Vázquez, N. </span> <span class="author-type"> </span> ; <span class="author-name" mtid="10027694"> <a href="/gui2/?type=authors&mode=browse&sel=10027694" target="_blank">Rona, G. ✉</a> </span> <span class="author-type"> </span> </div ><div class="title"><a href="/gui2/?mode=browse¶ms=publication;34487046" mtid="34487046" target="_blank">Visualizing Single-Stranded DNA Foci in the G1 Phase of the Cell Cycle</a></div> <div class="pub-info"> <span class="journal-title">JOVE-JOURNAL OF VISUALIZED EXPERIMENTS</span> <span class="journal-volume">2023</span> : <span class="journal-issue">202</span> <span class="page"> Paper: 65926 </span> <span class="year">(2023)</span> </div> <div class="pub-end"><div class="identifier-list"> <span class="identifiers"> <span class="id identifier oa_none" title="none"> <a style="color:black" title="10.3791/65926" target="_blank" href="https://doi.org/10.3791/65926"> DOI </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:black" title="85180673958" target="_blank" href="http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-85180673958"> Scopus </a> </span> </span> </div> <div class="short-pub-prop-list"> <span class="short-pub-mtid"> Közlemény:34487046 </span> <span class="status-holder"><span class="status-data status-APPROVED"> Nyilvános </span></span> <span class="pub-core">Forrás </span> <span class="pub-type">Folyóiratcikk (Szakcikk ) </span> <!-- && !record.category.scientific --> <span class="pub-category">Tudományos</span> </div> </div> </div><div class="JournalArticle Publication long-list">
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<span class="journal-issn">(<a target="_blank" href="https://portal.issn.org/resource/ISSN/1940-087X">1940-087X</a> <a target="_blank" href="https://portal.issn.org/resource/ISSN/1940-087X">1940-087X</a>)</span>:
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<pre class="comment" style="margin-top: 0; margin-bottom: 0;"><u>Megjegyzés</u>: Department of Biochemistry and Molecular Pharmacology, NYU Grossman School of Medicine, United States
The Laura and Isaac Perlmutter Cancer Center, NYU Langone Health, United States
Department of Cell Biology, NYU Grossman School of Medicine, United States
Howard Hughes Medical Institute, NYU Grossman School of Medicine, Un...</pre>
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