Ny doktorsavhandling

Från Australien kommer här en mastig doktorsavhandling utförd av f.d doktoranden Muthukuttige Madusha Nuwanthi Perera.Den numera titulerade Dr Perera måste sägas vara en stor anhängare av HoloMonitor.

I studien refereras till HoloMonitor inte mindre än 76 ggr !. Dr Perera får därmed anses vara en högst kvalificerad HoloMonitoranvändare. Han utgår förvisso från en stark HoloMonitor-bastion där instrumentet tidigare skördat en hel del segrar.Queensland University of Technology (QUT) där forskarna Derek Richard och Ken O'Byrne 2018 levererade en studie med rubriceringen :  

“We believe we’ve found the Holy Grail”

Bägge dessa forskare är vana HoloMonitoranvändare så Dr Perera har fått bästa utbildning från dem.

Men till studien benämnd :

Phytochemical analysis of two Australian native plants and their effects on chronic wound healing

Ur den tillhörande PDF-filen klipper jag in valda delar.

Abstract 

Chronic wounds are a significant burden to national health services worldwide and a potential reason for patients’ declining quality of life. It is estimated that the global market of wound management will reach $27.21 billion by 2028. Current costs of chronic wound management in Australia are over 3 billion AUD. Medicinal plants are widely used across a multitude of traditional medicine practices with minimal adverse effects. There is an emerging global trend of discovering effective and safe wound management therapies based on herbal plants. Indigenous Australians are known to use numerous medicinal plant species as traditional medicines. However, only a few have been scientifically investigated for their medicinal properties.

Från australiens växtrike har Dr Perera identifierat 2 växter som han nu bevisar har effekt på kroppens läkningsprocess .A. macrorrhizos och S. hillii. Med idogt användande av universitetets HoloMonitor levererar han nya fakta som är högst användbara.

Att kopiera alla hänvisningar till HoloMonitor låter sig inte göras. Menar, 76 st? Så jag tar enbart några få.

Instruments: Centrifuge Allegra X-15R (Beckman-Coulter, California, USA), POLAR star Omega plate reader (BMG Labtech, Mornington, Victoria, Australia), HoloMonitor M4 microscope (Phase Holographic Imaging AB, Lund, Sweden), QuantStudio 7 Flex real-time PCR system (Applied Biosystems, Thermo Fisher Scientific, Queensland, Australia). 

Software: Optima Data Analysis software version 2 (BMG Labtech Pty. Ltd., Ortenberg, Germany), HoloMonitor® App Suite software platform (version: 3.4.0) (Phase Holographic Imaging AB, Lund, Sweden), GraphPad Prism 8.4.3 software (GraphPad software, California, USA), QuantStudio 6 and 7 Flex real-time PCR system software v1.7.1 (Thermo Fisher Scientific, Queensland, Australia).  

4.3.4 Holographic live cell imaging assay

In order to identify potential morphological changes caused by the S. hillii methanol extract and its-derived compounds, HaCaT and HDFa cells were observed using the HoloMonitor®. 

HoloMonitor® live cell imaging uses a digital holography phase imaging technique to generate information based on phase shift of light on individual cells. It creates label-free images by dividing red laser light into a reference and an object beam. When the object beam passes through the cells, a phase delay is imprinted on the beam which later merged with the reference and recorded as a ‘holographic image’ .

Images were captured by HoloMonitor M4 microscope and analysed using HoloMonitor® App Suite software platform (version: 3.4.0). The HoloMonitor M4 is equipped with a 635 nm low laser beam with 0.2 mW/cm2 to prevent phototoxicity effects on the cells.

 

Figure 4.10. HoloMonitor® image analysis of S. hillii methanol extract-treated (3.9-62.5 µg/mL) HaCaT cells at 24 h, 48 h, and 72 h time points. Treatment with S. hillii methanol extract induced HaCaT cell proliferation over 72 h. Cells reached maximum confluence before the 72 h, hence, cells were shown to go through apoptosis and reduced cell volume. Untreated control: cells in RSM, Positive: cells in 10% FBS, Negative: cells in 10 mM thymidine. Each experiment performed in triplicate (n=3). Original magnification ×20. 

 

In order to resolve the discrepancy between results, a live cell imaging experiment was performed using a HoloMonitor M4 microscope. In contrast at 24 h, the A. macrorrhizos methanol extract did not exhibit any considerable effects on either cell line regardless of concentration tested. As such, no further testing was undertaken using the A. macrorrhizos methanol extract. 

HoloMonitor® live cell imaging system was used to confirm the effect of the S. hillii methanol extract as well as S. hillii methanol extract-derived compounds 3F1C1 and F6C3 (identified as shikimic acid and quercetin-3-O-β-D-glucuronide respectively in chapter 2) over 72 h. The concentration range selected was a four-fold concentration gradient from 3.9-62.5 µg/mL based on the significance of the results observed in both MTT and CyQUANT® assays. 

HoloMonitor® live cell imaging uses a digital holography phase imaging technique based on phase shift of light on individual cells, which can generate large amounts of information.

 

De övriga 65 hänvisningarna får ni leta upp själva.

Dr Perera avslutar doktorsavhandlingen med följande slutsatser :

 

In this project two bioactive compounds were isolated from the mature leaves of S. hillii: quercetin-3-O-β-D-glucuronide and shikimic acid. Both the primary methanol extract of S. hillii and isolated compounds exhibited significant antibacterial and anti-inflammatory effects, in addition to improving cell proliferation and migration under in vitro conditions. The isolated compounds were not cytotoxic to human-derived skin cells. Therefore, this project has scientifically confirmed the potential wound healing properties of S. hillii leaves, a remedy for chronic ulcers traditionally utilised by indigenous Australians. A potential S. hillii-based wound care product that modulates the important phases of wound healing would be beneficial to aged population as well as patients with chronic conditions such as diabetes, which in turn would reduce the burden to the healthcare sector.

 

Min kommentar

Det är nästan så man tycker att PHI borde skicka ett instrument till Dr Perera så han kan använda det gränslöst utan att behöva boka HoloMonitor-tid på universitet för vidare studier. Menar,med den hängivenheten och det idoga användande av HoloMonitor som svar på alla svåra ? som dök upp i studien får man nog säga att snubben vet hur man får ut max av instrumentet.

Vi gratulerar naturligtvis Dr Perera till sin väl värda titel.

 

                                           Mvh the99

 

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