Vol 3 Iss 3 by Peter G Mwitari
Ethnopharmacological relevance: Fungi have been used in traditional systems both as food and medi... more Ethnopharmacological relevance: Fungi have been used in traditional systems both as food and medicine. In China the mushrooms are widely used in Traditional Chinese Medicine (TCM). On one hand residue resulting from medicinal plants are treated as waste, yet the environment in which medicinal plants including fungi grow greatly influences the potency and quantities of bioactive secondary metabolites thereof. Since Mushrooms are widely consumed for immunomodulatory purposes especially for people suffering from tumors, this study sought to find out whether there is enhanced effect of mushroom polysaccharides extracts of Trametes robiniophila murr a Chinese fungus, grown on media supplemented with Chinese herbal drug residues both in vitro and in vivo as an immunotherapeutic remedy for cancer.
Methodology: We used Intestinal Epithelial Cells (IEC) known to produce IL-7 cytokine which works on the immune system and on the postulation that polysaccharide extracts work by up regulating IL-7. The polysaccharide extracts investigated were obtained from mushrooms grown on media supplemented with Chinese herbal residues. The Cell Counting Kit-8 (CCK-8) proliferation assay procedure was used to asses’ cell proliferation. Extract concentrations ranging from 0.00001 - 100μg/ml was used in the cell proliferation assay. IEC-6 cells grown in media supplemented with 100μg/ml mushroom polysaccharides extracts was used in vitro and RT-PCR technique employed to evaluate up-regulation of IL-7 in a time dependent manner ranging from 3 to 72 hours. The extract with best IL-7 expression was selected for further evaluation in vivo at 30mg/mice/day in mice infected with CT-26 colon cancer tumor cells. Cyclophosphamide at 20mg/mice/day was taken as the positive control drug. Flow cytometry was employed to evaluate the peripheral blood CD4+ and CD8+ lymphocytes. The extract was further tested in vitro on CT-26 cells (100 & 200 μg/ml), to explore direct inhibition.
Results: 1μg/ml was found to give optimum IEC-6 cell growth however; 100μg/ml was determined as the ED50 and used as the minimum evaluation concentration for IL-7 expression. The polysaccharide extracts obtained from mushrooms supplemented with the Chinese herb Gancao (Glycyrrhiza glabra L. var. glandulifera) residues were found to up-regulate IL-7 best. 6 hours exposure of cells to extract gave optimal IL7 expression of over 1.2 times at 100μg/ml compared to polysaccharide extracts from mushrooms grown on normal media. Extracts obtained at temperatures ranging between 70-90oC were found to be most ideal. The extract was found to increase the CD4+ and CD8+ lymphocytes in vivo in mice a reflection on IL-7 up regulation in the intestinal mucosa and further to inhibit CT-26 cell growth in a significant way at P≤0.05.
Conclusion: Polysaccharide extracts obtained from mushroom Trametes robiniophila murr grown on media supplemented with Chinese herb Gancao (G. glabra L. var. glandulifera) residues offers increased immunomodulatory effect as an alternative treatment to cancer cases. It can be employed as a prescription supplement for immunotherapy and immuno surveillance. The mechanism of action is probably stimulation of IEC-6 cells to produce increased IL-7 Cytokine responsible for resultant adaptive immunity and cancer immuno surveillance. The polysaccharide extract too has chemotherapeutic effect and is safe.
Key Words: Mushroom polysaccharides extracts, Trametes robiniophila murr, IL-7 Cytokine, CT-26 colon cancer tumor cells, CD4+ and CD8+ lymphocytes, immunity.
Papers by Peter G Mwitari
Withania somnifera, Warbugia ugandensis, Prunus africana and Plectrunthus barbatus are used tradi... more Withania somnifera, Warbugia ugandensis, Prunus africana and Plectrunthus barbatus are used traditionally in Kenya for treatment of microbial infections and cancer. Information on their use is available, but scientific data on their bioactivity, safety and mechanisms of action is still scanty. A study was conducted on the effect of organic extracts of these plants on both bacterial and fungal strains, and their mechanisms of action. Extracts were evaluated through the disc diffusion assay. Bacteria and yeast test strains were cultured on Mueller-Hinton agar and on Sabouraud dextrose agar for the filamentous fungi. A 0.5 McFarland standard suspension was prepared. Sterile paper discs 6 mm in diameter impregnated with 10 ml of the test extract (100 mg/ml) were aseptically placed onto the surface of the inoculated media. Chloramphenicol (30 mg) and fluconazole (25 mg) were used as standards. Discs impregnated with dissolution medium were used as controls. Activity of the extracts was e...
Journal of Pharmacognosy and Phytotherapy, Feb 28, 2018
Journal of Complementary and Alternative Medical Research
European Journal of Medicinal Plants
PloS one, 2018
Retroviral protease inhibitors (RPIs) such as lopinavir (LP) and saquinavir (SQ) are active again... more Retroviral protease inhibitors (RPIs) such as lopinavir (LP) and saquinavir (SQ) are active against Plasmodium parasites. However, the exact molecular target(s) for these RPIs in the Plasmodium parasites remains poorly understood. We hypothesised that LP and SQ suppress parasite growth through inhibition of aspartyl proteases. Using reverse genetics approach, we embarked on separately generating knockout (KO) parasite lines lacking Plasmepsin 4 (PM4), PM7, PM8, or DNA damage-inducible protein 1 (Ddi1) in the rodent malaria parasite Plasmodium berghei ANKA. We then tested the suppressive profiles of the LP/Ritonavir (LP/RT) and SQ/RT as well as antimalarials; Amodiaquine (AQ) and Piperaquine (PQ) against the KO parasites in the standard 4-day suppressive test. The Ddi1 gene proved refractory to deletion suggesting that the gene is essential for the growth of the asexual blood stage parasites. Our results revealed that deletion of PM4 significantly reduces normal parasite growth rate ...
Several studies nave been undertaken to deterrriine the karyotype of 5e50a171a species occurrmg i... more Several studies nave been undertaken to deterrriine the karyotype of 5e50a171a species occurrmg in tropical and sub-tropical reqions the world over. However, these studies have not been exhaustive. In the current research, a detailed karyotypic analysis of 13 .... sesoem» species found in southern Africa and collected for the first time by ICRAFwas undertaken. These species include; 5. sesoen 5. mecrenma. 5 rostrets. 5. c/oeresceos. 5 bispir/osa, 5 mi(Top/J.vlla, 5. 117aC()Wal7lal7;;1; 5: par..Yl,.v(:arp3., 5. sooserosoerms, 5. brevlpet1ul7cula, 5' tetreoters and 5. coerutescens Usmg HIe corwennonal chromosome techrnques, shoes were made, observed under the microscope and good spreads pnotormcroqraphed Trus studv conf irrneo HIe oprruon that HIe base number of HIe genus .... ")esbal7ia is X=6. A chromosome number of 2n=12 for 5. mscowemso« 5. pac!J..vcarp,3, 5. sooserosoerm« and 5. orevtoeamcot« however, was a new report. Two species namely; 5. tetrsoter» and 5. coerutsscen...
East African Medical Journal, 2008
Journal of Ethnopharmacology, 2007
Journal of Ethnopharmacology, 2011
Journal of Ethnopharmacology, 2008
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Vol 3 Iss 3 by Peter G Mwitari
Methodology: We used Intestinal Epithelial Cells (IEC) known to produce IL-7 cytokine which works on the immune system and on the postulation that polysaccharide extracts work by up regulating IL-7. The polysaccharide extracts investigated were obtained from mushrooms grown on media supplemented with Chinese herbal residues. The Cell Counting Kit-8 (CCK-8) proliferation assay procedure was used to asses’ cell proliferation. Extract concentrations ranging from 0.00001 - 100μg/ml was used in the cell proliferation assay. IEC-6 cells grown in media supplemented with 100μg/ml mushroom polysaccharides extracts was used in vitro and RT-PCR technique employed to evaluate up-regulation of IL-7 in a time dependent manner ranging from 3 to 72 hours. The extract with best IL-7 expression was selected for further evaluation in vivo at 30mg/mice/day in mice infected with CT-26 colon cancer tumor cells. Cyclophosphamide at 20mg/mice/day was taken as the positive control drug. Flow cytometry was employed to evaluate the peripheral blood CD4+ and CD8+ lymphocytes. The extract was further tested in vitro on CT-26 cells (100 & 200 μg/ml), to explore direct inhibition.
Results: 1μg/ml was found to give optimum IEC-6 cell growth however; 100μg/ml was determined as the ED50 and used as the minimum evaluation concentration for IL-7 expression. The polysaccharide extracts obtained from mushrooms supplemented with the Chinese herb Gancao (Glycyrrhiza glabra L. var. glandulifera) residues were found to up-regulate IL-7 best. 6 hours exposure of cells to extract gave optimal IL7 expression of over 1.2 times at 100μg/ml compared to polysaccharide extracts from mushrooms grown on normal media. Extracts obtained at temperatures ranging between 70-90oC were found to be most ideal. The extract was found to increase the CD4+ and CD8+ lymphocytes in vivo in mice a reflection on IL-7 up regulation in the intestinal mucosa and further to inhibit CT-26 cell growth in a significant way at P≤0.05.
Conclusion: Polysaccharide extracts obtained from mushroom Trametes robiniophila murr grown on media supplemented with Chinese herb Gancao (G. glabra L. var. glandulifera) residues offers increased immunomodulatory effect as an alternative treatment to cancer cases. It can be employed as a prescription supplement for immunotherapy and immuno surveillance. The mechanism of action is probably stimulation of IEC-6 cells to produce increased IL-7 Cytokine responsible for resultant adaptive immunity and cancer immuno surveillance. The polysaccharide extract too has chemotherapeutic effect and is safe.
Key Words: Mushroom polysaccharides extracts, Trametes robiniophila murr, IL-7 Cytokine, CT-26 colon cancer tumor cells, CD4+ and CD8+ lymphocytes, immunity.
Papers by Peter G Mwitari
Methodology: We used Intestinal Epithelial Cells (IEC) known to produce IL-7 cytokine which works on the immune system and on the postulation that polysaccharide extracts work by up regulating IL-7. The polysaccharide extracts investigated were obtained from mushrooms grown on media supplemented with Chinese herbal residues. The Cell Counting Kit-8 (CCK-8) proliferation assay procedure was used to asses’ cell proliferation. Extract concentrations ranging from 0.00001 - 100μg/ml was used in the cell proliferation assay. IEC-6 cells grown in media supplemented with 100μg/ml mushroom polysaccharides extracts was used in vitro and RT-PCR technique employed to evaluate up-regulation of IL-7 in a time dependent manner ranging from 3 to 72 hours. The extract with best IL-7 expression was selected for further evaluation in vivo at 30mg/mice/day in mice infected with CT-26 colon cancer tumor cells. Cyclophosphamide at 20mg/mice/day was taken as the positive control drug. Flow cytometry was employed to evaluate the peripheral blood CD4+ and CD8+ lymphocytes. The extract was further tested in vitro on CT-26 cells (100 & 200 μg/ml), to explore direct inhibition.
Results: 1μg/ml was found to give optimum IEC-6 cell growth however; 100μg/ml was determined as the ED50 and used as the minimum evaluation concentration for IL-7 expression. The polysaccharide extracts obtained from mushrooms supplemented with the Chinese herb Gancao (Glycyrrhiza glabra L. var. glandulifera) residues were found to up-regulate IL-7 best. 6 hours exposure of cells to extract gave optimal IL7 expression of over 1.2 times at 100μg/ml compared to polysaccharide extracts from mushrooms grown on normal media. Extracts obtained at temperatures ranging between 70-90oC were found to be most ideal. The extract was found to increase the CD4+ and CD8+ lymphocytes in vivo in mice a reflection on IL-7 up regulation in the intestinal mucosa and further to inhibit CT-26 cell growth in a significant way at P≤0.05.
Conclusion: Polysaccharide extracts obtained from mushroom Trametes robiniophila murr grown on media supplemented with Chinese herb Gancao (G. glabra L. var. glandulifera) residues offers increased immunomodulatory effect as an alternative treatment to cancer cases. It can be employed as a prescription supplement for immunotherapy and immuno surveillance. The mechanism of action is probably stimulation of IEC-6 cells to produce increased IL-7 Cytokine responsible for resultant adaptive immunity and cancer immuno surveillance. The polysaccharide extract too has chemotherapeutic effect and is safe.
Key Words: Mushroom polysaccharides extracts, Trametes robiniophila murr, IL-7 Cytokine, CT-26 colon cancer tumor cells, CD4+ and CD8+ lymphocytes, immunity.