Cancer Therapy: GcMAF, Nagalase and Macrophage

Body's defence system
Our body contains cell stores which help us to generate cells which fight cancer. These stores or nodes are present throughout the body.

Macrophage & phagocyte locations
Macrophages and phagocytes are produced by the skin, gut and intestinal Peyer's patches, lungs, blood, bone marrow, connective tissue, lymphoid tissue, spleen, and thymus.

Types of phagocytes

1. Skin - macrophages, resident Langerhans cells, dendritic cells, mast cells
2. Gut and intestinal Peyer's patches - macrophages
3. Lungs - macrophages, monocyes, mast cells, dendritic cells
4. Blood - neutrophils, monocytes
5. Bone marrow - macrophages, monocytes, sinusoidal cells, lining cells
6. Connective tissue - macrophages, monocytes, dendritic cells, histiocytes
7. Lymphoid tissue - macrophages, monocytes, dendritic cells
8. Spleen - macrophages, monocytes, sinusoidal cells
9. Thymus - macrophages, monocytes

Peyer's patches
The nodes in the intestine are most important. These nodes are called Peyer's patches. They are found beneath the basement membrane of the intestinal mucosa. They are a source of macrophages and immunoglobulin A (Ig A) in response to contents of the food a person consumes.

Invaders that invade the human body

1. Viruses (viral infection)
2. Cancer

Viral infection
(keywords: viral enzymes (nagalases), macrophages, immunity)
When a batch of virus launch an attack on the body, the viruses produce an array of enzymes that weaken the macrophages, rendering them ineffective in fighting off the viruses. This is described as a weakened immune system because the body fails to defend itself and fails to fight off the invading viruses.

Cancer
Cancer cells also produce nagalases which act on macrophages, rendering them weak and inefficient in protecting the body against invading cancer cells. With a weakened immune system, the cancer is able to grow and metastasise.

GcMAF-nagalase loop

1. GcMAF (contains Gal-GalNAc)
2. Nagalase

GcMAF
GcMAF is Gc Protein-derived Macrophage Activating Factor. GcMAF is a glycosylated protein bound to vitamin D. There are three forms (subtypes) of GcMAF. They differ in the number of sugar and sialic acid residues in the side chain. The sugar found  in GcMAF are glycosamines.

Structure of Gc protein
The Gc protein contains a threonine (Thr) residue, to which is attached a disaccharide side chain containing galactose (Gal) and Galactosamine (GalNAc) and sialic acid (SA), or simply, Gal-GalNAC-SA. Gal-GalNAC is a cancer biomarker for colon cancer (carcinogenesis). There are three forms of Gc which can be converted to GcMAF. They are Gc1f, Gc1s and Gc2.

(Gc1f):  -aa-aa-aa-aa-Thr-aa-aa-
                                    |
                        Gal-GalNAc
                                    |
                                  SA

Gc1s: -aa-aa-aa-aa-Thr-aa-aa-
                                 |
           (SA)-Gal-GalNAc
                                |
                              SA

Gc2: -aa-aa-aa-aa-Thr-aa-aa-
                               |
                   Gal-GalNAc


Nagalase
Nagalase if one of the viral enzymes. There are two types of nagalases - ie exo- and endo-nagalases. They act at different sies (loci) on GcMAF.

Pathway
When virus or cancer attacks the body, they produce nagalases, which act on macrophages and inactivate them. GcMAF on the other hand, acts opposite of nagalase, ie, they promote macrophage activation. Activated macrophages can transform into natural killer cells (NK cells) which can actively fight a viral infection or cancer.

Production of GcMAF
GcMAF is produced by a laboratory in Japan. There are two productions of GcMAF, ie first and second generations.

1. First generation GcMAF (1991-2010)
2. Second generation GcMAF (2011-present)

First generation GcMAF

This GcMAF was less concentrated, unstable at room temperature and had a short half-life.

Second generation GcMAF

This GcMAF was more concentrated, more stable and can last two weeks without refrigeration and up to a year with refrigeration. This is the preferred form for use.

Administration routes of GcMAF
GcMAF can be administered via four routes, ie, skin, muscle, intestine and lungs.

1. Skin (subcutaneous, SC)
2. Muscle (intramuscular, IM)
3. Intestine (oral)
4. Lungs (inhalation via nebulizer)

Conversion of Gc1f to GcMAF

1. B cells contain lyso-PC inducible beta-galactosidase. This enzyme removes galactose from the Gal-GalNAc side chain.
2. T cells contain sialidase. This enzyme removes sialic acid from the GalNAc-SA side chain.

GcMAF: -aa-aa-aa-aa-Thr-aa-aa-

                                      |

                                 GalNAc

Macrophage activation and transformation

GcMAF acts on dormant macrophages to activate them. Activated macrophages transform to NK cells.

Is GcMAF therapy effective?

YES, in the 99 patients who obtained successful treatment.

NO, in one patient in which treatment failed.

Failure rate is 1 in 100 or 1% ineffectiveness, which is negligible.

External links
https://www.immunotherapy-cancer-and-chronic-disease.com/frequently-asked-questions-about-second-generation-gcmaf/

http://gcmaf.timsmithmd.com/book/chapter/53/

https://www.advancedrejuvenationinstitute.com/cancer/3-immunotherapy/cancer-research-gcmaf.html

https://healingoracle.ch/2017/09/06/tutorial-how-to-inject-gcmaf/

https://www.researchgate.net/figure/Plasticity-of-macrophages-in-tumor-microenvironment-and-pathogenesis-of-HCC_fig2_275041931

https://www.immunopaedia.org.za/clinical-cases/infectious-diseases/a-case-of-decreased-joint-function-fever-and-rash/

http://cancerres.aacrjournals.org/content/76/3/513/F1

https://healingoracle.ch/2017/09/06/tutorial-how-to-inject-gcmaf/

https://cancer-cures-plus.com/persecuted-heroes/how-gcmaf-works/new-cancer-fighter-gcmaf-update/

https://thetruthaboutcancer.com/gcmaf/

http://www.brainimmune.com/endogenous-catecholamines-in-immune-cells-discovery-functions-and-clinical-potential-as-pharmacotherapeutic-targets-3/

https://immuno-oncologynews.com/2015/06/09/pd-1-protects-t-cells-burning/

https://www.ncbi.nlm.nih.gov/pubmed/8839767

Biomarker of Cisplatin-Induced Nephrotoxicity

INTRODUCTION

CISPLATIN

Cisplatin is used in chemotherapy but with nephrotoxicity as the most common side effect. Thus, cancer patients treated with cisplatin can suffer from acute renal failure (ARF) which is fatal.

Cisplatin is a chemotherapy drug and contains platinum that is complexed. It is an anti-cancer drug. It is also known as platinol or Platin. It has a half life of 300-100 hours and is excreted via the kidneys. Cisplatin is used to induce ARF in mice. Cisplatin inhibits fatty acid oxidation, and fatty acids accumulate in affected cells. Cisplatin induces nephrotoxicity (toxic to the renal cells). The platinum complex reacts in vivo, binding to and causing crosslinking of DNA (DNA cease to function), which ultimately triggers apoptosis (programmed cell death).  Cisplatin causes necrosis. It is used for treatment of solid malignancies. It is used to treat various types of cancers, including sarcomas, some carcinomas (e.g. small cell lung cancer, and ovarian cancer), lymphomas, and germ cell tumors. Cisplatin is particularly effective against testicular cancer; the cure rate was improved from 10% to 85%. In addition, Cisplatin is used in Auger therapy (to breakup herpes genes with ionising radiation from platinum).

From Wikipedia

PPARalpha

Fibrate treatment (with PPARalpha) ameliorates (relieves, improves) renal function by preventing the inhibition of fatty acid oxidation and proximal tubule cell death. Peroxisome proliferator-activated receptor-alpha (PPARalpha) is a nuclear receptor protein. It serves to overcome the effects of cisplatin in renal cells.

Function

PPAR-alpha is a transcription factor and a major regulator of lipid metabolism in the liver. PPAR-alpha is activated under conditions of energy deprivation and is necessary for the process of ketogenesis, a key adaptive response to prolonged fasting. Activation of PPAR-alpha promotes uptake, utilization, and catabolism of fatty acids by upregulation of genes involved in fatty acid transport, fatty binding and activation, and peroxisomal and mitochondrial fatty acid β-oxidation. PPAR-alpha is primarily activated through ligand binding.  Endogenous ligands include fatty acids and various fatty acid-derived compounds. Synthetic ligands include the fibrate drugs, which are used to treat hyperlipidemia. Insecticides, herbicides, plasticizers, and organic solvents also bind to PPAR-alpha, and are collectively referred to as peroxisome proliferators.

Tissue distribution

Expression of PPAR-alpha is highest in tissues that oxidize fatty acids at a rapid rate. In rodents, highest mRNA expression levels of PPAR-alpha are found in liver and brown adipose tissue, followed by heart and kidney. Lower PPAR-alpha expression levels are found in small and large intestine, skeletal muscle and adrenal gland. Human PPAR-alpha seems to be expressed more equally among various tissues, with high expression in liver, intestine, heart, and kidney.

From Wikipedia

METABOLIC ABNORMALITIES IN ARF

There are several systemic metabolic abnormalities which occur prior to and coexist with the development of ARF. They are responsible for the accumulation of free fatty acids in ischaemic ARF. They include:

1. glucose intolerance 
2. insulin resistance
3. accumulation of fatty acids (accumulation of free fatty acids and toxic long chain fatty acid metabolites in freshly isolated proximal tubules subjected to hypoxic injury, and in kidney tissue of animals subjected to ischemia/reperfusion injury)
4. activation of intracellular calcium-independent phospholipase A2
5. inhibition of mitochondrial fatty acid oxidation 

CREATININE

In the past, creatinine was often measured to determine creatinine clearance and hence assess renal function. However, creatinine levels change slowly in blood and alterations in creatinine often go undetected. So we research on ARF, cisplatin and PPARalpha in live mice models instead.


METHODOLOGY

1. Lab animals: Mice

ARF is studied in live laboratory mice models.

Urine samples of mice treated with single injection of cisplatin (20 mg/kg body weight) were collected for 3 days and analyzed by 1H—nuclear magnetic resonance (NMR) spectroscopy.

2. Test drugs

Two drugs are used - cisplatin and PPARalpha.

3. Investigations

There are many ways to study the effects of cisplatin and PPARalpha on kidneys:

1. Biochemical analyses (chemicals in urine; urine metabolites)
2. Electron microscopy (tissue specific changes; cytopathology)

Biochemical analyses

Biochemical analysis of endogenous metabolites was performed in serum, urine, and kidney tissue.

Principal component analysis demonstrated the presence of glucose, amino acids, and trichloacetic acid cycle (TCA cycle) metabolites in the urine after 48 h of cisplatin administration. These metabolic alterations precede changes in serum creatinine.

 Urine metabolite concentrations are measured by 1H NMR spectroscopy:

Alanine --> amino acid (Ala)
Glucose --> glucosuria (sweet urine)
Lactate --> organic acid; end product from anaerobic glycolysis
Leucine --> amino acid (Leu)
Methionine --> amino acid (Met)
1-Methylnicotinamide --> methylated NADH produced by glycolysis
2-Oxoglutarate --> TCA cycle intermediate
Proline --> amino acid (Pro)
Pyruvate --> organic acid; end product from aerobic glycolysis
Trimethylamine --> internal standard for 1H NMR
Tyrosine --> amino acid (Tyr)
Valine --> amino acid (Val)

The urine metabolite concentrations are normalised to the concentration of creatinine in urine. Values are expressed as mg metabolite/mg creatinine.

Electron microscopy

Electron microscopic studies were carried out to examine the effects of PPARalpha ligand and cisplatin.

Electron microscopic analysis confirmed the protective effect of the fibrate on preventing cisplatin-mediated necrosis of the S3 segment of the proximal tubule.

RESULTS

1. Biochemical studies confirmed the presence of glucosuria, but also demonstrated the accumulation of nonesterified fatty acids, and triglycerides in serum, urine, and kidney tissue, in spite of increased levels of plasma insulin. 
2. These metabolic alterations were ameliorated by the use of PPARalpha ligand. 
3. When cisplatin is administered into mice, there are metabolites found in urine - they appear with time.

CONCLUSION

The study shows that cisplatin-induced a unique NMR metabolic profile in urine of mice that developed ARF, and confirmed the protective effect of a fibrate class of PPARalpha ligands.

BIOMARKER PROPOSAL

The researchers proposed that the injury-induced metabolic profile may be used as a biomarker of cisplatin-induced nephrotoxicity.


External links:
http://www.nature.com/ki/journal/v69/n12/full/5000433a.html
http://en.wikipedia.org/wiki/Cisplatin
http://en.wikipedia.org/wiki/Peroxisome_proliferator-activated_receptor_alpha
http://en.wikipedia.org/wiki/Auger_therapy

Risk factors for cancer

Dr. Epstein cites the 2008-2009 Annual Report of the President’s Cancer Panel, released in April 2010, includes a “Summary of Environmental and Occupational Links with Cancer.” This report documents “strong” evidence on cancer risks from exposures to 15 individual or groups of carcinogens, such as talc powder, ethylene oxide, and dioxane. The report also documents “suspected” evidence from exposure to the larger range of risks from exposure to about 40 other individual or groups of carcinogens.

Source: World Wire

Further work:

1) Make a list of cancer risks  & carcinogens.

2) Make a questionnaire & distribute (print or online).

3) Collect feedback & analyse.

4) Report.

Cancer

Cancer rate increases in our society. A cure for cancer for an afflicted person is the sarang angkut-angkut (wasp hive) in his own house, and the pokok semalu (forget-me-not plant). For any disease or condition that involves swelling, the pokok semalu is the cure as it will reduce the swelling.

Take the pokok semalu, air zamzam and sarang angkut-angkut, and pound to a paste. Read Ayat al-Quran. Apply the paste to boils, swellings, etc. It will feel cool. It takes the heat away.

Read Al-Fatihah, Ayat Kursi & Surah Al-Baqarah Ayat 22.

Source: tv9, Tanyalah Ustaz (Ustaz Sharhan Shafie), Penyakit & Penawarnya, 25 Sept 2011, 7.40am 

Cancer patient fund raising

Raising money for a cancer patient but she died.

http://www.youtube.com/watch?v=RGpZ9bchCJw&feature=related

How beneficial is the citrus?

What studies say
Scientific studies indicate compounds in citrus, including lemon, have real potential as anti-cancer agents. Studies have indicated that citrus limonoids do have potential as anti-cancer agents.

Grey areas
However, it is not yet clear exactly HOW effective citrus will ultimately prove to be in preventing or fighting against cancer in humans. It is PREMATURE and INACCURATE to claim that lemon is a "proven remedy against cancers of all types." Nor, at this point, can it be said that lemon is a viable alternative to traditional treatments such as chemotherapy. But lemon, like other kinds of citrus is likely to be a healthy addition to your diet and may even reduce the risk of cancer. However, these findings do not give validity to the exaggerated and unsupported claims made wrt lemons so far. To be useful, health advice needs to be valid, accurate and be supported by credible medical sources. Sending on spurious health information is unlikely to be beneficial.

Research results we have so far
A number of studies have indicated that compounds found in citrus (including lemon) may be effective as anti-cancer agents, at least for certain types of cancer. A December 2004 Science Daily article reports:

Research by Texas Agriculture Experiment Station scientists has shown that citrus compounds called limonoids targeted and stopped neuroblastoma cells in the lab. They now hope to learn the reasons for the stop-action behavior and eventually try the citrus concoction in humans. [......]

Harris explained that flavonoids and limonoids – nutrient-packed pigments that give color and taste to fruit – may work against cancer in any of three ways: prevent it from forming, slow the growth of existing cancer, or kill cancer cells.

"The limonoids, which differ structurally from flavonoids, seem to do all three," he said of tests in his lab by one of Patil's graduate students, Shibu Poulose, who also worked in Harris' College Station lab. Their work emphasized the compounds' ability to kill the existing neuroblastoma cells with the rationale that if the method and time limonoids take to obliterate the cancer could be found, perhaps scientists could exploit it to help cure the disease.

A May 2000 report about the potential of citrus limonoids as anticancer agents explains:

The experimental results described above indicate that citrus limonoids may provide substantial anticancer actions. The compounds have been shown to be free of toxic effects in animal models so potential exists for use of limonoids against human cancer in either the natural fruit , in citrus fortified with limonoids, or in purified forms of specific limonoids . Although the initial studies are very promising , they have been conducted primarily with in vitro cell culture and animal models. Thus , research is needed to determine whether the limonoids may be useful in preventing or treating cancer in humans .

And a report on the medicinal use of citrus published on the University of Florida EDIS website notes:

Citrus flavonoids have potential antioxidant (prevents aging), anti-cancer, antiviral, anti-inflammatory activities, effects on capillarity, and cholesterol-lowering ability. The principal carotenoids in pink grapefruit are lycopene and beta-carotene. Lycopene-containing fruits and vegetables have been shown to contribute to a significant reduction in prostate and mammary cancer risk. Recent studies have further shown that limonoids inhibit the development of cancer in laboratory animals and in human breast cancer cells as well as reducing cholesterol. Researchers have also suggested that, if ingested, limonoids may not be absorbed in the large intestine, and therefore could be distributed throughout the body, with beneficial effects.

 Source: http://www.hoax-slayer.com/lemon-cancer-cells.shtml