Chemotherapies Tested

we send you the results from the analysis made about a patient (patient's name) suffering from breast carcinoma stage IV . The sample that was sent to us for analysis was a sample of 25ml of whole blood that contained EDTA-Ca as anti-coangulant, all packed with water ice. In our laboratory we made the following:

    We isolated the malignant cells using Oncoquick with a membrane that isolates malignant cells from normal cells after centrifugation and positive selection using epithelial cell marker negative selection using anti-CD45 particles .
    Then we developed cell cultures in a fetal calf serum media and at the same time we developed colony cultures in soft agar. In each culture of the well plate we added a chemotherapeutic substance that is used in clinical application. Then we developed those cultures and we harvested a sample every 24 hours for 6 days and made the following assays.
    There was made an isolation of the genomic DNA using the kit Invisorb of INVITEK.
    We isolated mRNA using the mRNA Magprep blood isolation kit of NOVAGEN.
    We traced the mRNA and the genes of MDR1 ( multi drug resistant 1 ), MRP and LRP using the technique of Northern Blot .(resistance in drugs used in chemotherapies)
    We tracked the mRNA and the gene of topoisomerase I and II a & b using the technique of Northern Blot . ( sensitivity in cytostatic inhibitors of topoisomerase )
    We tracked the quantity of the mRNA of the tubulin using the RT-PCR.( sensitivity in cytostatics of the kind of taxanes and the products of the alkaloids of Vinca )
    We defined the activity of the enzyme complex of the gloutathion-S- transferees (GST kit of NOVAgen) . ( resistance in drugs used in chemotherapies- especially in platinum compounds)
    We defined the DNA methyl transferase which is a target of the alkyliating factors (products of platinum , cyclophosphamide and the products of it)
    We defined the mRNA of the thymidylate synthetase ( TS ) and the DHFR . (sensitivity in 5-FU, capecitabine and methotrexate )
    We defined the mRNA of the reductase of 5-CMP (sensitivity in gemcitabin)
    We defined the receptors of the MMP and the receptors of laminin (invasive ability of the tumor)
    We defined the expression of protein p27 that is responsible for cell arrest in GO stage.
    We defined the VEGF ( neoangiogenetic factor ) and the induction of the apoptotic pathway using ONCOGENE kit from NOVAgen.
    We defined the ability of acting of the nucleous protein kinases which are a target of the carbazin compounds .
    We defined the overexpression of TGFa and TGFb factors as targets for suramin sulfate.
    We defined the overexression of somatostatin receptor (SS-R), of COX-2 and 5-LOX , of c-erb-B2 (Her/Neu2) , c-erb-Bl, and androgen estrogen and progesterone receptors.

1. From the whole neoplasmic population we have an expression of MDRl in a percentage of 50% over control sample .(positive in the check of resistance )
2. From the whole neoplasmic population we have an expression of MDRl in a percentage of 50% over control sample .(positive in the check of resistance )
3. The activity of gammaGC is stable in the low limits (no resistance to platinum compounds)
4. The activity of CES1 and CES2 is in normal range (no resistance to camptothecin compounds)
5. The concentration of pi80 is in high range
6. Increased activity of the laminin and the MMP ( increased invasive ability)
7. There is partial sensitivity in taxanes (paclitaxel, docetaxel) and partial sensitivity noticed in alkaloids of Vinca.
8. Minimal sensitivity noticed in 5FC, in 5-FU, in UFT , in FUdR in capecitabine, in raltitrexed, in methotrexate , in pemetrexed but there is great sensitivity in gemcitabine.
9. Increased sensitivity in alkyliating factors (especially in cyclophosphamide and melphalan).
10. There is great overexpression of TGF b (40% over control) , of NFkBeta (35% over control) and EGF-r (50%,<70%) growth factors and supression of expression of isoforms of IKB (a, d, e) (40% below control) .
11. It appears to have no sensitivity in the inhibitors of topoisomerase II a and II b .
12. There is great sensitivity in the inhibitors of topoisomerase I (especially in topotecan).
13. There is no overexpression of SS-r receptor and there is no overexpression of COX-2 mRNA , of 5-LOX mRNA but there is great overexpression c-erb-B2 (50% over control), of c-erb-Bl (55% over control), of estrogen receptor mRNA (45% over control) and of progesterone receptor mRNA (10% over control).
14. We notice great neoangiogenetic ability (overexpression of VEGF-R - 60% over control sample).
15. Finally, there is no sensitivity in dacarbazine .
16. We notice that taurolidin cannot induce the apoptosis to the malignant cells (in IV route dosage).
17. We notice that taurolidin can induce the apoptosis to the malignant cells (in intraperitoneal route dosage)

CONCLUSION:

    The specific tumor appears to have resisting populations because of the MDR1 overexpression that can be reversed by the use of verapamil combined with imidazole compounds (ketoconazole).
    The neoplasmatic cells have the greatest sensitivity in the alkyliating agents cvclophosphamide and melphalan, in the inhibitor of topoisomerase I topotecan and in the antagonist gemcitabine.
    Also you can use: erlotinib as inhibitor of EGF-r , bortezomib as inhibitor of proteasome over-activity and indirectly the transcriptional activity of NFKB, 5-azacytidine as inhibitor of DNA hypermethylation,anti- estrogen as inhibitors of estrogen receptor overexpression, transtuzubab as inhibitor of c-erb-B2 (Her/neu2), normal p53 (advexin) as inducer of apoptosis, and bevacizumab as inhibitor of angiogenesis.

-------------------------------------------------------------------------------------------------------------------------------------

Thalidomide

we send you the results from the analysis made about a patient (patient's name) suffering from breast carcinoma stage IV. The sample that was sent to us for analysis was a sample of 25ml of whole blood that contained EDTA-Ca as anti-coangulant, all packed with water ice . In our laboratory we made the following:

    We isolated the malignant cells using Oncoquick with a membrane that isolates malignant cells from normal cells after centrifugation and positive selection using epithelial cell marker negative selection using anti-CD45 particles.
    We develop two (2) different cultures from malignant cells (one with thalidomide[th+] in the culture media -in concentration AUC- and one without thalidomide[th-]) from the blood sample of patient above.
    From the culture that include thalidomide [th+] to the media in the culture with malignant cells, we measure the activity of caspase 3 and cytochrome c.
    From both cultures we make compared analysis of VEGF, PDGF, FGF and MMPs inhibition rate.


The results are presented below:

Malignant cell cultures compared analysis

Conclusion: We notice that the thalidomide can inhibit the neovascularization and it can induce the apoptosis to the cancer cell becoming from the patient above, but it cannot inhibit the invasion activity of the cancer cells.

-------------------------------------------------------------------------------------------------------------------------------------

Biological Modifiers (Alternative Holistic Substances)

we send you the results from the analysis made about a patient (patient's name) suffering from breast carcinoma stage IV. The sample that was sent to us for analysis was a sample of 25ml of whole blood that contained EDTA-Ca as anti-coangulant . packed with water ice . In our laboratory we made the following:

    We isolated the malignant cells using Oncoquick with a membrane that isolates malignant cells from normal cells . Then we centrifuged at 350g for 10 min and we collected the supernatant with the malignant cells. Then we proceed to isolation of malignant cells from mononuclear cells by negative selection .
    Then we developed forty-one cell cultures in a fetal calf serum media . In each culture of the well plate we added a biological modifier substance (H2O2, ascorbic acid , carnivora , misteltoe, quercetin , indol-3-carbinol , c-statin , ukrain , polyMVA, co enzyme Q10, essiac tea, modified citrus pectin, IP6 , pacreatic enzymes, salvestrol, Uncaria Tomentosa, carctol, noni juice, annonaceous acetogenins, caesium chloride, reolysin. amygdalin-B17-, artesunate, maitake, lycopene, curcumin, green tee extract, melatonin, ellagic acid, L-methionine, N-acetyl-cystein, Niacin (Vit.B3), L- carnithine, Vitamin E (tocopherol), superoxide dismutase (SOD), selenium, aloe vera, IFNa2, clim basical tea, clim tea drc, tap-V) that is used in clinical application. Then we developed those cultures and we harvested a sample every 24 hours and made the following assays.
    In the culture that it contains all substance we measure the apoptotic ability using the oncogen apoptosis kit
    In the culture that it contains the carnivora we measure the inhibition of tyrosin kinase catalytic ability from growth factors receptor (EGF-r, IGF-r,) and the production of cytokines.
    In the culture that it contains the Ukrain we measure the inhibition of tyrosin kinase catalytic ability from growth factors receptor (EGF-r, IGF-r,) and the production of cytokines PMBC
    In the culture that contains quercetin we measure the inhibition of EGF and IGF .
    In the culture that contains indol-3-carbinol we measure the inhibition of VEGF and FGF and PDGF
    In the culture that it contains the misteltoe we measure the inhibition of tyrosin kinase catalytic ability from growth factors receptor (EGF-r, IGF-r,) and the production of cytokines and the increase of PMBC
    In the culture that it contains the H2O2 we measure viability of the culture in 4 days of treatment.
    In the culture that it contains the ascorbic acid we measure the catalytic activity of GSH and GSSG (redox reaction) and the induction of cytochrome C (apoptosis).
    In the culture that it contains the PolyMVA we measure the catalytic activity of GSH and GSSG (redox reaction) and the induction of cytochrome C (apoptosis).
    In the culture that it contains the artesunate we measure the catalytic activity of GSH and GSSG (redox reaction for free radical since artesunate bind free radicals with iron molecule ) , the inhibition of VEGF , FGF and PDGF (since it act to the angiogenesis cascade reactions) and the induction of cytochrome C (apoptosis).
    In the culture that it contains the artesunate we measure the catalytic activity of GSH and GSSG (redox reaction for free radical since artesunate bind free radicals with iron molecule ) , the inhibition of VEGF , FGF and PDGF (since it act to the angiogenesis cascade reactions) and the induction of cytochrome C (apoptosis).

RESULTS:
1. We notice that in culture that contains the ascorbic acid we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by 50% .
2. We notice that in culture that contains the PolyMVA we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by 45% .
3. We notice that in culture that contains carnivora we have inhibition of EGF-r by less than 5% and for IGF-r by 5% and we notice no increase of cytokine production 5% .
4. We notice that in the culture that contains quercetin we have inhibition of EGF by 55% and IGF by 50%
5. We notice that in the culture that contains indol-3-carbinol we have inhibition of VEGF by 60% , of FGF by 60% , and PDGF by 55%
6. We notice that in culture that contains misteltoe we have inhibition of EGF-r by 50% and for IGF-r by 45% and we notice increase of cytokine production by 70%, and there is increase of PMBC by 70%.
7. We notice that in culture that contains the c-statin we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by 50%.
8. We notice that in culture that contains Ukrain we have inhibition of EGF-r by less than 5% and for IGF-r by 5% and we notice no increase of cytokine production, and there is no increase of PMBC.
9. We notice that in culture that contains the H2O2 we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable .
10. We notice that in culture that contains the co enzyme Q10 we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable.
11. We notice that in culture that contains the essiac tea we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable .
12. We notice that in culture that contains the modified citrus pectin we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable .
13. We notice that in culture that contains the IP6 we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable .
14. We notice that in culture that contains the pacreatic enzymes we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable .
15. We notice that in culture that contains the salvestrol we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable.
16. We notice that in culture that contains the uncaria tomentosa we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable.
17. We notice that in culture that contains the caesium chloride we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
18. We notice that in culture that contains the carctol we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable.
19. We notice that in culture that contains the noni juice we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable.
20. We notice that in culture that contains the annonaceous acetogenins we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable.
21. We notice that in culture that contains the reolysin we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by less than 5% and the viability of the culture remain stable.
22. We notice that in culture that contains the amygdalin-B 17- we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
23. We notice that in culture that contains maitake we have inhibition of EGF-r by less than 5% and for IGF-r by 5% and we notice no increase of cytokine production , and there is no increase of PMBC.
24. We notice that in culture that contains the curcumin (turmeric) we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
25. We notice that in culture that contains the lykopene we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
26. We notice that in culture that contains the green tea extract we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable
27. We notice that in culture that contains artesunate , there is inhibition of redox reaction and increase of intracellular free radicals, there is increase of cytochrome c (apoptosis) by 65% and the inhibition rate of VEGF is 55%, of FGF is 50% and of PDGF is 45%.
28. We notice that in culture that contains the melatonin we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
29. We notice that in culture that contains the ellagic acid we have increase of the cascade of caspase (especially 3 and 9) and cytochrom-c by 60% and the viability of the culture reduced by 40%.
30. We notice that in culture that contains the L-methionine we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
31. We notice that in culture that contains the N-acetyl-cystein we have noincrease of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
32. We notice that in culture that contains the niacin we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
33. We notice that in culture that contains the L-eamithine we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
34. We notice that in culture that contains the vitamin E we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
35. We notice that in culture that contains the superoxide dismutase we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
36. We notice that in culture that contains the aloe vera extract we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
37. We notice that in culture that contains selenium we have inhibition of EGF-r by less than 5% and for IGF-r by 5% and we notice no increase of cytokine production , and there is no increase of PBMC and NK .
38. We notice that in culture that contains IFNa2 we have inhibition of EGF-r by less than 5% and for IGF-r by 5% and we notice no increase of cytokine production , and there is no increase of PMBC .
39. We notice that in culture that contains tap-V we have inhibition of EGF-r by 25% and for IGF-r by 20% and we notice increase of cytokine production by 95% , and there is increase of PMBC by 80% .
40. We notice that in culture that contains the clim basical tea we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.
41. We notice that in culture that contains the clim tea drc we have no increase of the cascade of caspase (especially 3 and 9) and cytochrom-c and the viability of the culture remain stable.

CONCLUSION: It seems that this specific population of malignant cell have greater sensitivity in quercetin, in artesunate, in Poly-MVA, in indol - 3-carbinol, in mistletoe, in c-statin, ellagic acid, tap-V and in ascorbic acid and less in hydrogen peroxide (H2O2), co enzyme Q10, essiac tea, modified citrus pectin, IP6 , N-acetyl-cystein, pacreatic enzymes, salvestrol, in carnivora, in L-carnithine, in L-methionine, vitamin E (tocopherol), maitake, in ukrain, caesium chloride, in IFNa2. , superoxide dismutase, amygdalin-(B17), in curcumin (turmeric), uncaria tomentosa (samento), in melatonin, in selenium, carctol, noni juice, niacin, aloe vera extract, annonaceous acetogenins (paw paw), reolysin (reovirus), lykopene, clim basical tea, clim teat drc and in green tea extract.

-------------------------------------------------------------------------------------------------------------------------------------