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Methods and Protocols, Volume 7, Issue 6

December 2024 - 22 articles

Cover Story: With the growing interest in malaria-derived extracellular vesicles (EVs), standardizing the isolation protocols is essential for ensuring reproducibility and enabling inter-study comparisons. This article provides a detailed, reproducible differential centrifugation (DC) protocol for isolating EVs from Plasmodium falciparum-infected red blood cell cultures at various asexual blood stages. The method yields two EV subtypes from high-parasitemia, synchronized cultures, characterized by the Western blot analysis of human and parasite proteins, and identifies the potential protein markers for malaria EV subpopulations. Researchers can use this protocol to facilitate downstream applications for further characterisation, such as proteomics and functional studies, advancing our understanding of malaria biology. View this paper
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Articles (22)

  • Article
  • Open Access
1,736 Views
9 Pages

Evaluation of Protocols for DNA Extraction from Individual Culex pipiens to Assess Pyrethroid Resistance Using Genotyping Real-Time Polymerase Chain Reaction

  • Ilaria Congiu,
  • Elisa Cugini,
  • Daniele Smedile,
  • Federico Romiti,
  • Manuela Iurescia,
  • Valentina Donati,
  • Claudio De Liberato and
  • Antonio Battisti

23 December 2024

Culex pipiens is a major vector of pathogens, including West Nile and Usutu viruses, that poses a significant public health risk. Monitoring pyrethroid resistance in mosquito populations is essential for effective vector control. This study aims to e...

  • Article
  • Open Access
1,785 Views
17 Pages

21 December 2024

Recommendations released by the CDC in 2023 address the need to demonstrate that the RNA genome of positive-strand RNA viruses is inactivated in addition to viral particles. This recommendation is in response to the similarities between host mRNA and...

  • Protocol
  • Open Access
1 Citations
1,790 Views
12 Pages

Establishment of Stable Knockdown of MACC1 Oncogene in Patient-Derived Ovarian Cancer Organoids

  • Sophia Hierlmayer,
  • Liliia Hladchenko,
  • Juliane Reichenbach,
  • Christoph Klein,
  • Sven Mahner,
  • Fabian Trillsch,
  • Mirjana Kessler and
  • Anca Chelariu-Raicu

20 December 2024

High-grade serous ovarian cancer (HGSOC) remains the most lethal gynecological malignancy, and there is still an unmet medical need to deepen basic research on its origins and mechanisms of progression. Patient-derived organoids of high-grade serous...

  • Article
  • Open Access
1,427 Views
10 Pages

20 December 2024

The accurate measurement of spatiotemporal parameters, such as step length and step frequency, is crucial for analyzing running and sprinting performance. Traditional methods like video analysis and force platforms are either time consuming or limite...

  • Article
  • Open Access
1 Citations
1,797 Views
13 Pages

20 December 2024

An electrochemical investigation of 1,2- and 1,4-dihydroxybenzenes was carried out with platinum macro- and microelectrodes using square wave and cyclic voltammetry techniques. Furthermore, the effect of the two solvents—acetic acid and ethyl a...

  • Article
  • Open Access
3,067 Views
13 Pages

Proof of Concept Study: Comparison of Semi-Automated RNA Isolation Methods from Archived Formalin-Fixed, Paraffin-Embedded Tissues with Clinical Routine RNA Isolation Methods

  • Patrick Hannibal Dalsbo Petersen,
  • Jaslin Pallikkunnath James,
  • Lene Buhl Riis,
  • Claus Kim Høgdall and
  • Estrid Vilma Høgdall

19 December 2024

High-quality RNA is crucial in clinical diagnostics and precision medicine. Formalin-fixed and paraffin-embedded (FFPE) tissues pose a challenge due to nucleic acid fragmentation and crosslinking. In this pilot study, various commercially available t...

  • Protocol
  • Open Access
2,545 Views
26 Pages

The Multifunctional Catalytic Hemoglobin from Amphitrite ornata: Protocols on Isolation, Taxonomic Identification, Protein Extraction, Purification, and Characterization

  • Anna L. Husted,
  • Victoria R. Sutton,
  • Lauren A. Presnar,
  • R. Kevin Blackburn,
  • Joseph L. Staton,
  • Stephen A. Borgianini and
  • Edward L. D’Antonio

11 December 2024

The multifunctional catalytic hemoglobin from the terebellid polychaete Amphitrite ornata, also named dehaloperoxidase (AoDHP), utilizes the typical oxygen transport function in addition to four observed activities involved in substrate oxidation. Th...

  • Protocol
  • Open Access
1 Citations
2,568 Views
13 Pages

Liver Cancer Neuroscience: Regulating Liver Tumors via Selective Hepatic Vagotomy

  • Kylynda C. Bauer,
  • Shadin Ghabra,
  • Chi Ma,
  • Lee Chedester and
  • Tim F. Greten

11 December 2024

Both the prevalence and mortality of liver cancers continue to rise. Early surgical interventions, including liver transplantation or resection, remain the only curative treatment. Nerves in the periphery influence tumor growth within visceral organs...

  • Study Protocol
  • Open Access
1 Citations
1,907 Views
13 Pages

10 December 2024

Background: About 287,000 women died globally during their pregnancy journey in 2020, yet most of these deaths could have been prevented. In Uganda, studies show that using Community Health Worker (CHW) visits to households with a pregnant woman can...

  • Article
  • Open Access
2 Citations
1,477 Views
14 Pages

2 December 2024

Cyclic peptides have higher stability and better properties as therapeutic agents than their linear peptide analogues. Consequently, intramolecular click chemistry is becoming an increasingly popular method for the synthesis of cyclic peptides from t...

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Methods Protoc. - ISSN 2409-9279