scholarly journals Synthesis, characterisation, and application of chamomile gold nanoparticles in molecular diagnostics: a new component for PCR kits

2019 ◽  
Vol 9 (6) ◽  
pp. 4635-4641 ◽  
Keyword(s):  
Gold Nanoparticles ◽  
Nucleic Acids ◽  
Low Cost ◽  
Gc Content ◽  
Nucleic Acid Amplification ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain ◽  
Successful Amplification

Various strategies have been suggested for successful amplification of the hard-to-amplify nucleic acids such as the inclusion of various chemical or biological materials in the in vitro nucleic acid amplification reactions, particularly polymerase chain reaction (PCR), and adjustment of the cycling programs. Although much efforts have been madefor improvements in the enhancement of polymerase chain reactions of high GC content nucleic acids, still significant challenges remain. In this study, the effects of citrate-coated AuNP and chamomile extract-coated AuNP (p-AuNP) on amplification of three genes with significant different GC percentage were evaluated. Owing to the enormous potential of gold nanoparticles in the enhancement of the PCR reactions, we showed the promising and consistent findings on the application of very dilute biocompatible chamomile-gold nanoparticles as safe and low-cost nanomaterials for molecular amplification of GC-rich DNA samples. We hypothesized that green AuNPs, which have different surface chemistry from cit-AuNPs, not only do not interfere with the PCR reactants but also are capable of enhancing PCR reactions. These results, for the first time, confirm the potential of using the green gold nanoparticles in the heat-assisted enzymatic in vitro reactions, suggesting Chamomile gold nanoparticles as reliable component of any PCR kits.

scholarly journals Influence of mesenchymal stem cell-derived extracellular vesicles in vitro and their role in ageing

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Juan Fafián-Labora ◽  
Miriam Morente-López ◽  
María José Sánchez-Dopico ◽  
Onno J. Arntz ◽  
Fons A. J. van de Loo ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Extracellular Vesicles ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Transmission Electron ◽  
Pluripotency Markers ◽  
Polymerase Chain ◽  
Mtor Signalling

Abstract Introduction This study assessed whether mesenchymal stem cell (MSC)-derived extracellular vesicles influenced ageing and pluripotency markers in cell cultures where they are added. Methods MSC-derived extracellular vesicles from old and young rat bone marrows were isolated by ultracentrifugation and were characterised by western blotting, nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). They were added to young and old MSC cultures. Real-time quantitative reverse transcription polymerase chain reactions and western blot analysis were performed to check the markers of ageing (vinculin and lamin A), pluripotency markers (Nanog and Oct4) and components of the mTOR signalling pathway (Rictor, Raptor, AKT and mTOR) in these cell populations. Subsequently, microRNA (miR)-188-3p expression was transiently inhibited in young MSCs to demonstrate the influence of mTOR2 on MSC ageing. Results Incubation with young MSC-derived extracellular vesicles decreased the levels of ageing markers and components of the mTOR pathway and increased the pluripotency markers from old MSC populations. By contrast, incubation of young MSCs with old MSC-derived extracellular vesicles generated the reverse effects. Inhibition of miR-188-3p expression in young MSCs produced extracellular vesicles that when incubated with old MSCs produced an increase in the levels of Rictor, as well as a decrease of phosphor-AKT, as indicated by a significant decrease in beta-galactosidase staining. Conclusions MSC-derived extracellular vesicles affected the behaviour of MSC cultures, based on their composition, which could be modified in vitro. These experiments represented the basis for the development of new therapies against ageing-associated diseases using MSC-derived extracellular vesicles.

scholarly journals Nucleic acid amplification by a transparent graphene Visual-PCR chip and a disposable thermocycler

2019 ◽  
Author(s):  
Guozhi Zhu ◽  
Miao Qiao
Keyword(s):  
Nucleic Acids ◽  
Color Change ◽  
Point Of Care ◽  
Low Cost ◽  
Nucleic Acid Amplification ◽  
Driving Voltage ◽  
Conductive Films ◽  
Cooling Fan ◽  
Resource Limited ◽  
Polymerase Chain

ABSTRACTPolymerase chain reaction (PCR) is a method widely used to amplify trace amount of nucleic acids. It needs a process of thermocycling (repeated alternation of temperature). Traditional thermocycler relies on bulk size of metal block to achieve thermocycling, which results in high cost and the lack of portability. Here, a PCR chip made of graphene Transparent Conductive Films (TCFs) was employed. The thermocycling of the chip was fulfilled by a temperature programed microcontroller and a cooling fan under a low driving voltage (12V). A 35 cycles PCR was accomplished within 13 minutes using the chip and the thermocycler. The transparency of the graphene PCR chip enables the PCR reaction to be visually monitored by naked eye for a color change. The PCR chip and the thermocycler have a low cost at $2.5 and $6 respectively, and thus are feasible for Point-of-care testing (POCT) of nucleic acids in a disposable manner. The whole platform makes it possible to perform a low-cost testing of nucleic acids for varieties of purposes outside of laboratories or at resource limited locations.

scholarly journals Investigation of Lethal Concurrent Outbreak of Chlamydiosis and Pigeon Circovirus in a Zoo

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1654
Author(s):  
Wei-Tao Chen ◽  
Chin-Ann Teng ◽  
Cheng-Hsin Shih ◽  
Wei-Hsiang Huang ◽  
Yi-Fan Jiang ◽  
...  
Keyword(s):  
Disease Outbreaks ◽  
Chlamydia Psittaci ◽  
Bursa Of Fabricius ◽  
Chain Reactions ◽  
Renal Epithelium ◽  
Polymerase Chain Reactions ◽  
Transmission Electron ◽  
Intracytoplasmic Inclusion Bodies ◽  
Polymerase Chain ◽  
Streptopelia Orientalis

During the spring, an outbreak of sudden death involving 58 birds occurred in a zoo. Histopathological examinations revealed variable numbers of intracytoplasmic basophilic microorganisms in the macrophages, hepatocytes, and renal epithelium of most birds, along with occasional botryoid intracytoplasmic inclusion bodies within histiocytes in the bursa of Fabricius. Based on the results of histopathological examinations, immunohistochemical staining, transmission electron microscopy, and polymerase chain reactions, genotype B Chlamydia psittaci infection concurrent with pigeon circovirus (PiCV) was diagnosed. A retrospective survey, including two years before the outbreak and the outbreak year, of C. psittaci and PiCV infections of dead birds in the aviaries, revealed that the outbreak was an independent episode. The findings of this study indicate that concurrent infection with C. psittaci and PiCV might lead to lethal outbreaks of chlamydiosis, particularly Streptopelia orientalis. In addition, persistently monitoring both pathogens and identifying potential PiCV carriers or transmitters might also help prevent lethal disease outbreaks.

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