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Methods and Protocols, Volume 6, Issue 4

2023 August - 14 articles

Cover Story: Loss of lysosomal membrane integrity affects cellular functions and can induce cell death via the release of lysosomal hydrolases. Several drugs used clinically have the ability to destabilize the lysosome, which can be utilized as a treatment strategy. The expanding interest for lysosomal function has emphasized the need for assays that follow lysosomal damage and detect minor damage that does not inevitably cause cell death. The weak base acridine orange accumulates in the acidic environment of lysosomes but is released upon lysosomal damage. In this protocol, we present a high-throughput microplate reader-based method to follow destabilization of the lysosomal membrane in real time using acridine orange. This protocol can easily be adopted for patient samples since the number of cells per sample is low and the time for analysis is short. View this paper
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Articles (14)

  • Protocol
  • Open Access
26 Citations
8,649 Views
11 Pages

Real-Time Monitoring of Lysosomal Membrane Permeabilization Using Acridine Orange

  • Ida Eriksson,
  • Linda Vainikka,
  • Hans Lennart Persson and
  • Karin Öllinger

Loss of lysosomal membrane integrity results in leakage of lysosomal hydrolases to the cytosol which might harm cell function and induce cell death. Destabilization of lysosomes often precede apoptotic or necrotic cell death and occur during both phy...

  • Article
  • Open Access
32 Citations
8,734 Views
12 Pages

Recent advances in genomic technologies have enabled more in-depth study of the oral microbiome. In this study, we compared the amplicons generated by primers targeting different sites of the 16S rRNA gene found in the Human Oral Microbiome Database...

  • Article
  • Open Access
2 Citations
4,606 Views
16 Pages

Application of a Customised Franz-Type Cell Coupled with HPTLC to Monitor the Timed Release of Bioactive Components in Complex Honey Matrices

  • Md Lokman Hossain,
  • Minh Nguyen,
  • Leah Benington,
  • Lee Yong Lim,
  • Katherine Hammer,
  • Dhanushka Hettiarachchi and
  • Cornelia Locher

The aim of this study was to assess the release profile of components in five different honeys (a New Zealand Manuka and two Western Australian honeys, a Jarrah honey and a Coastal Peppermint honey) and their corresponding honey-loaded gel formulatio...

  • Article
  • Open Access
14 Citations
4,189 Views
13 Pages

Ultrasound-Assisted Anthocyanins Extraction from Pigmented Corn: Optimization Using Response Surface Methodology

  • Annisa Nurkhasanah,
  • Titouan Fardad,
  • Ceferino Carrera,
  • Widiastuti Setyaningsih and
  • Miguel Palma

This study aimed to determine the optimal UAE conditions for extracting anthocyanins from pigmented corn using the Box–Behnken design (BBD). Six anthocyanins were identified in the samples and were used as response variables to evaluate the eff...

  • Protocol
  • Open Access
3 Citations
2,348 Views
13 Pages

Geriatric assessment (GA) is fundamental to optimising cancer care in older adults, yet implementing comprehensive GA tools in real-world clinical settings remains a challenge. This study aims to assess the feasibility and acceptability of integratin...

  • Protocol
  • Open Access
1 Citations
2,103 Views
7 Pages

(1) Background: Oncological demolitive–reconstructive surgeries in the head and neck region cause significant stress on patients’ biohumoural, cardiac, and vascular systems, leading to disturbances in macrocirculatory and microcirculatory...

  • Protocol
  • Open Access
2,725 Views
9 Pages

Ex Vivo Perfusion of Porcine Pancreas and Liver Sourced from Commercial Abattoirs after Circulatory Death as a Research Resource: A Methodological Study

  • Zainab L. Rai,
  • Morenike Magbagbeola,
  • Katie Doyle,
  • Lukas Lindenroth,
  • George Dwyer,
  • Amir Gander,
  • Agostino Stilli,
  • Danail Stoyanov and
  • Brian R. Davidson

Background: Machine perfusion (MP) is increasingly used for human transplant organ preservation. The use of MP for research purposes is another opportunity for this technology. The porcine pancreas and liver are similar in anatomical size and functio...

  • Article
  • Open Access
6 Citations
2,199 Views
9 Pages

Assessment of SARS-CoV-2 (COVID-19) Clinical Mouthwash Protocol and Prevalence of the Oral Pathogen Scardovia wiggsiae: A Pilot Study of Antibacterial Effects

  • Melika Shayegh,
  • Chase Sorenson,
  • Jackson Downey,
  • Summer Lin,
  • Yuxin Jiang,
  • Praneeti Sodhi,
  • Victoria Sullivan,
  • Katherine M. Howard and
  • Karl Kingsley

One protocol in healthcare facilities and dental offices due to the COVID-19 pandemic for reducing the amount of detectable oral SARS-CoV-2 has been gargling with mouthwash for 60 s. This protocol lasts longer than the daily routine for most patients...

  • Article
  • Open Access
2 Citations
2,765 Views
11 Pages

Development of a Wound-Healing Protocol for In Vitro Evaluation of Urothelial Cell Growth

  • Christopher Foster,
  • Todd Jensen,
  • Christine Finck and
  • Courtney K. Rowe

Urethral healing is plagued by strictures, impacting quality of life and medical costs. Various growth factors (GFs) have shown promise as therapeutic approaches to improve healing, but there is no protocol for in vitro comparison between GFs. This s...

  • Brief Report
  • Open Access
85 Citations
16,886 Views
6 Pages

As an essential characterization, size distribution is an important indicator for the synthesis, optimization, and application of nanoparticles. Electron microscopes such as transmission electron microscopes (TEMs) are commonly utilized to collect si...

  • Protocol
  • Open Access
10 Citations
4,703 Views
8 Pages

A New and Profitable Protocol to DNA Extraction in Limnospira maxima

  • Yirlis Yadeth Pineda-Rodriguez,
  • Marcelo F. Pompelli,
  • Alfredo Jarma-Orozco,
  • Novisel Veitía Rodríguez and
  • Luis Alfonso Rodriguez-Paez

Limnospira maxima is a remarkable organism showing great potential as a versatile and sustainable food source, offering a powerful solution to address the pressing issues of malnutrition and undernourishment worldwide. L. maxima contains high amounts...

  • Protocol
  • Open Access
1 Citations
3,182 Views
10 Pages

Proposed Protocol for Field Testing of Endurance Fitness of Young Labrador Retrievers

  • Ella-Erika Söderlund,
  • Heikki Kyröläinen,
  • Outi M. Laitinen-Vapaavuori and
  • Heli K. Hyytiäinen

The number of dogs and, with it, dog sports are growing in popularity, and the training of dogs begins at an early age. Although fitness testing is an imperative part of purposeful training and sports, to our knowledge, no objective field tests are a...

  • Review
  • Open Access
22 Citations
4,604 Views
15 Pages

The detection of toxic heavy metal ions, especially cadmium (Cd), lead (Pb), zinc (Zn), and copper (Cu), is a global problem due to ongoing pollution incidents and continuous anthropogenic and industrial activities. Therefore, it is important to deve...

  • Technical Note
  • Open Access
1 Citations
1,944 Views
6 Pages

Optimized Protocol for Preservation of Human Platelet Samples for Fluorometric Polyphosphate Quantification

  • Tomoyuki Kawase,
  • Katsuya Suzuki,
  • Masami Kamimura,
  • Tomoharu Mochizuki and
  • Takashi Ushiki

Platelet polyphosphate (polyP) can be conveniently quantified by exploiting a recent methodological breakthrough using 4′,6-diamidino-2-phenylindole (DAPI). However, the preservation of these biological samples has not yet been standardized. In...

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Methods Protoc. - ISSN 2409-9279