Happy palm Sunday world. Tonight in the magnificent chapel i enjoyed a choir and string concert. The chapel was full. They made room for ne. I had my infusion pole. I got some palms but i missed mass.
The conference being in holy week well easter is about life after death. New life. Maybe life after cancer. Not a mention about root cause analysis of cancer and the immune failures that lead to our struggle.
Dear god grant me courage strength and wisdom.
PERSONAL MEDICAL
PH 6.8 glucose 6.4 basal temp 36.7 for ketones 1.0 continue improvements in all markers indicating a reduction of inflammation and healing.
Im finishing off my 4th bottle of taurolidine. Its Sunday night and i have 6 hours driving to munich for the conference. Im glad it starts 11am.
CONFERENCE FOREPLAY
Dear god . Help me to manage the drive safely and arrive ready to learn.
The storm of knowledge and ideas coming from these scientists i made. God grant me wisdom in abundance to find my way. I've already done what these GIFTED SCIENTISTS present as rat AND mouse success.
Its a very exciting conference program.
http://www.ecco-org.eu/Events/ITOC3/Searchable-Programme#anchorScpr
Hypoxic/acidic tumour microenvironment could be linked to inflammatory/suppressive cells and may induce an aggressive phenotype in human hepatocellular carcinoma.
GCMAF POSTER
Introduction
Vitamin D binding protein when deglycosylated is a naturally potent macrophage stimulatory factor known as Group Component Macrophage Activation Factor (GcMAF). Data has been presented that demonstrate GcMAF as reducing tumour burden in several clinical approaches. GcMAF has stimulated Raw 264.7 murine macrophages in vitro that phagacytose MCF-7 cells.In this study we demonstrate the direct effect of three proteins GcMAF a recombinant GcMAF and a truncated GcMAF peptide on the ability to affect the growth of Raw 264.7 and MCF7 cells. In addition data on the ability of the three proteins to stimulate the RAW 264.7 macrophages to phagacytose the MCF7 cells.
Materials and Methods
Cell lines purchased from the ECACC Cultures Public Health England UK.
RAW264.7: Murine macrophages
MCF7: Human Breast Adenocarcinoma
Macrophages were stimulated with either GcMAF, Recombinant GcMAF or synthetic polypeptide
Cell viability assay
Viability was evaluated by reduction of WST-8 as an index of cell activity. The cell line was treated with GcMAF 8 80 or 800 pM. After incubation 10µl of WST-8 is added. The resulting OD45nm was directly measured.
Cells were plated into wells at a known density in starvation medium. After incubation they were treated with the GcMAF species at 8 80 or 800 pM. After incubation the cell suspension was counted.
Video photography
MCF7 cells were seeded prior to the addition of the GcMAF activated RAW macrophages. The well cultures were observed and an image selected. An initial image was taken and a time-lapse film initiated. A frame was taken every 3 min.
Results
Cell viability
The results were compared to non-activated cells. The proliferation assay used the RAW macrophages when compared to the fully glycosylated GcGlobulin the native deglycoslyated GcMAF caused a 25% decrease in proliferation, the synthetic polypeptide had a 26% reduction whilst the recombinant GcMAF was 36%. These results confirm that the recombinant GcMAF and the synthetic polypeptide convey the same effect on the cells lines as the native GcMAF protein.
Time-lapse video
Selected frame shots from the time-lapse of cells activated with the different GcMAF species highlight the macrophage capability to phagacytose the MCF7 cells.
Discussion
It has been shown that GcMAF activates RAW macrophages to phagocytose and destroy MCF-7 cells. This demonstrated that alternative GcMAF species acts upon human and murine macrophages in the same way to destroy cancer cells.
Conclusions
Here the activating effect on macrophages can be achieved using native, recombinant and synthetic GcMAF polypeptide. The effect on cell line proliferation is similar and the ability to stimulate murine macrophages to destroy human breast carcinoma cells is shown. The recombinant and synthetic approach alleviates the issues of the native protein in terms of the safety of the material moving forwards.
