EDIBLEPIECE

Research

Research at ediblepiece

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Oncology

Cell culturing.
Generation of polyplexes by self assembly method.
Development and optimisation of a robust procedure for localisation of nanoparticles in the cell.
Protein isolation, identification and quantification.
Identification of peptide and nucleic acids using the bioinformatic tools.
Isolation and identification of protein biomarker in cell sample by immunolabelling.
Endocytosis pathways in live and fixed cells.
Mechanism of delivery of molecules in cells.

Read about our research on cancer here

Infection and immunology

Signaling study; RNA isolation and quantification using qPCR.
Protein extraction and quantification using western blotting.
Immunoflourescence and protein localisation using confocal laser scanning microscopy.
Usage of software such as qPCR analysing programme, Biorad microplate reader , ZEISS examiner, IMAGEJ image analyser, Odyssey infrared imaging system, Kodak imaging system etc.
Bacterial cultivation and infection; multiplicity of infection (MOI).

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Nanotechnology

Development of sensor based on RNA aptamer.
Establishment of protocols for gene therapy.
Establishment of protocols for analysis of cellular endocytic vesicles.
confocal microscopy and immunofluorescence.
Colocolisation of nanoplexes with caveolae, ascertaining the relationship and their location in the caveolae.
Confocal microscopy image acquisition and analysis.
Transfection with luciferace plasmid.

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Pharmacology

Incontinence

Read about our research on incontinence (urinary and fecal) here

Chemical pathology

Development of optimal and robust Invivo microscopic protocol for tissue analysis.
Development and optimisation of robust Immunohistological techniques for tissue imaging.
Development and optimisation of analytical protocol for quantification of biochemical body fluids.
Detection and quantification of malondialdehyde (MDA) in animal tissue.
Optimisation of protein isolation, purification and identification in animal organs.
Determination and quantification of body lipid; HDL, LDL, free cholesterol level.
Urine analysis; Optimisation of protocol for detection and quantification of bence jones protein.
Optimisation of a robust protocol for detection and quantification of urine protein, sugar and blood.General body electrolyte quantification.
Establishing protocols for detection and correlation of level of blood bicarbonate and chloride with disease state.
Detection, quantification and correlation of level of blood sugar in diabetes patients; Fasting blood sugar (FBS) and Random blood sugar.
Development, optimisation and application of microscopy in microbial analysis.Generation and optimisation of body fluid analysis protocol .
Optimisation and application of immunological techniques in microbial detection and quantification.
Application of immunoprecipitation method in microbial detection.
Detection and quantification of microbes by colorimetry method.
Protocol establishment for blood type verification and genotyping.

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Papers

Int J Biol Macromol. 2012 Dec;51(5):1043-51. doi: 10.1016/j.ijbiomac.2012.08.016. Epub 2012 Aug 27.

Cellular uptake of DNA-chitosan nanoparticles: the role of clathrin- and caveolae-mediated pathways.Garaiova Z1, Strand SP, Reitan NK, Lélu S, Størset SØ, Berg K, Malmo J, Folasire O, Bjørkøy A, Davies Cde L.

Author information

Abstract

The success of gene therapy depends on efficient delivery of DNA and requires a vector. A promising non-viral vector is chitosan. We tailored chitosan to optimize it for transfection by synthesizing self-branched and trisaccharide-substituted chitosan oligomers (SBTCO), which show superior transfection efficacy compared with linear chitosan (LCO). The aim of the work was to compare the cellular uptake and endocytic pathways of polyplexes formed by LCO and SBTCO. Both polyplexes were taken up by the majority of the cells, but the uptake of LCO was lower than SBTCO polyplexes. LCO polyplexes were internalized through both clathrin-dependent and clathrin-independent pathways, whereas SBTCO polyplexes were primarily taken up by clathrin-independent endocytosis. The different level of cellular uptake and the distinct endocytic pathways, may explain the difference in transfection efficacy. This was supported by the observation that photochemical internalization increased the transfection by LCO polyplexes considerably, whereas no effect on transfection was found for SBTCO polyplexes.

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Abstracts

Davies C deL, Garaiova Z, Reitan NK, Bjørkøy A, Folasire O, Størset S, Berg K, Strand SP:

DNA-chitosan nanoparticles in gene delivery: Endocytotic pathways and intracellular trafficking.

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Oladayo S Folasire, Kylie Mills, Donna Sellers, Russ Chess-williams.

Characterisation of relaxatory transmitters in the porcine internal anal sphincter

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Oladayo S Folasire, Donna Sellers, Russ Chess-Williams

Characterisation of contractile responses to alpha1 agonists in the porcine urethral circular smooth muscle.

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Biotechnology

Extraction of protein from cell, determination of total protein in cell lysate by bicinchoninic acid protein assay (BCA).
Resolution of total protein on gel by electrophoresis in order to detect and quantify specific protein of interest such as LC3I, LC3II and Keap1.
Using microbe "mycobacterium avium" to stimulate autophagy.
Measuring autophagy by confocal microscopy based on immunoflurescence.
Cultivation of cells such as Hela and macrophages inorder to develop an autophagy read out protocol.
Usage of Bio-rad plate reader in quantifying the total protein in the cell lysate.
Usage of Odyssey imager and Kodak imager in detecting the protein bands as resolve on the gel after blotting.
Usage of Kodak analyser and Odyssey analyser to quantify the protein band.
Staining blot for protein of interest and housekeeping protein such as GAPDH.Normalised the protein bands with housekeeping gene such as GAPDH.
Preparing cell for imaging by fixing, permeabilising, blocking and labeling with antibody.
Transfection of macrophages with GFP-LC3 or mcherry-GFP-LC3 conjugate plasmid using gene juice kit.
Transforming bacterial with GFP-LC3 and GFP-mcherry-LC3 conjugates plasmid.

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