Identification and characterization of main allergic proteins in cooked Wolf herring fish

Our aim in this study was to identify and characterize allergic proteins in cooked Wolf herring fish. We heated the crude extract alternatively at 50, 60, 70, 80, 90, and 100°C for one hour and results were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Also, proteins were immunoblotted with fish-sensitive patients' sera. The major allergenic proteins were identified via mass spectrometry. These allergenic proteins were then purified by anion exchange chromatography and the IgE-immunoreactivity of the fractions was compared with the crude extracts via disk enzyme-linked immunosorbent assay (ELISA). SDS-PAGE of the crude extract showed more than 15 distinct protein bands. Five of these proteins, with apparent molecular weights of 12, 18, 24, 38, and 51 kDa, were only observed in the 100°C heated extract. Immunoblotting of the heated extract revealed that the 12 and 51 kDa proteins were IgE-immunoreactive with 88 percent of fish-sensitive patient sera while the 24 and 38 kDa proteins reacted with 33.3 and 55.5 percent of fish-sensitive patient sera, respectively. Mass spectrometry of the 12, 38, and 51 kDa proteins revealed that all three were parvalbumin oligomers. Disk ELISA results showed that 20 of 25 and 14 of 25 fish-allergic patients' sera were IgE-reactive with purified oligomeric parvalbumin-coated and crude extract-coated disks, respectively. Parvalbumin and its oligomers are the main allergenic molecules in cooked fish. Therefore, an enriched or purified fraction containing this protein could be a useful source of allergen for applications in ELISA-based immunoassays and could discriminate fish-allergic patients who can tolerate cooked fish from those who cannot. © Copyright Autumn 2016, Iran J Allergy Asthma Immunol. All rights reserved

Cross-linking gold nanoparticles aggregation method based on localised surface plasmon resonance for quantitative detection of miR-155

MiR-155 plays a critical role in the formation of cancers and other diseases. In this study, the authors aimed to design and fabricate a biosensor based on cross-linking gold nanoparticles (AuNPs) aggregation for the detection and quantification of miR-155. Also, they intended to compare this method with SYBR Green real-time polymerase chain reaction (PCR). Primers for real-time PCR, and two thiolated capture probes for biosensor, complementary with miR-155, were designed. Citrate capped AuNPs (18.7 ± 3.6 nm) were synthesised and thiolated capture probes immobilised to AuNPs. The various concentrations of synthetic miR-155 were measured by this biosensor and real-time PCR method. Colorimetric changes were studied, and the calibration curves were plotted. Results showed the detection limit of 10 nM for the fabricated biosensor and real-time PCR. Also, eye detection using colour showed the weaker detection limit (1 µM), for this biosensor. MiR-133b as the non-complementary target could not cause a change in both colour and UV�visible spectrum. The increase in hydrodynamic diameter and negative zeta potential of AuNPs after the addition of probes verified the biosensor accurately fabricated. This fabricated biosensor could detect miR-155 simpler and faster than previous methods. © The Institution of Engineering and Technology 2017

miR-219 overexpressing oligodendrocyte progenitor cells for treating compression spinal cord injury

Oligodendrocyte progenitor cells (OPCs) transplantation has been considered a promising treatment for spinal cord injury, according to previous studies. Recent research shed light on the importance of microRNA 219 (miR-219) in oligodendrocyte development, so here miR-219-overexpressing OPCs (miR-219 OPCs) were transplanted in animal models of spinal cord injury to evaluate the impact of miR-219 on oligodendrocyte differentiation and functional recovery in vivo. Our findings demonstrate that transplanted cells were distributed in the tissue sections and contributed to reducing the size of cavity in the injury site. Interestingly, miR-219 promoted OPC differentiation into mature oligodendrocyte expressing MBP in vivo whereas in absence of miR-219, less number of cells differentiated into mature oligodendrocytes. An eight week evaluation using the Basso Beattie Bresnahan (BBB) locomotor test confirmed improvement in functional recovery of hind limbs. Overall, this study demonstrated that miR-219 promoted differentiation and maturation of OPCs after transplantation and can be used in cell therapy of spinal cord injury. © 2021, The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature

Cross-linking gold nanoparticles aggregation method based on localised surface plasmon resonance for quantitative detection of miR-155

MiR-155 plays a critical role in the formation of cancers and other diseases. In this study, the authors aimed to design and fabricate a biosensor based on cross-linking gold nanoparticles (AuNPs) aggregation for the detection and quantification of miR-155. Also, they intended to compare this method with SYBR Green real-time polymerase chain reaction (PCR). Primers for real-time PCR, and two thiolated capture probes for biosensor, complementary with miR-155, were designed. Citrate capped AuNPs (18.7 ± 3.6 nm) were synthesised and thiolated capture probes immobilised to AuNPs. The various concentrations of synthetic miR-155 were measured by this biosensor and real-time PCR method. Colorimetric changes were studied, and the calibration curves were plotted. Results showed the detection limit of 10 nM for the fabricated biosensor and real-time PCR. Also, eye detection using colour showed the weaker detection limit (1 µM), for this biosensor. MiR-133b as the non-complementary target could not cause a change in both colour and UV�visible spectrum. The increase in hydrodynamic diameter and negative zeta potential of AuNPs after the addition of probes verified the biosensor accurately fabricated. This fabricated biosensor could detect miR-155 simpler and faster than previous methods. © The Institution of Engineering and Technology 2017