[Study on anti-cancer effect of plant extract ACA-1 on gastric adenocarcinoma cell line]

Apoptosis provides solutions for the treatment of cancers and its induction is used as a strategy for preparing drugs that destroy pre-neoplastic cells. Many plant compounds have anti-tumoral activity. ACA-1 plant product is an aqueous extract and has been used in traditional medicine in Iran and has cytotoxicity effect on melanoma cancer cells. In this study, anti-cancer effect of ACA-1 plant product on gastric adenocarcinoma cells was evaluated and the mechanism of its action was studied. Cytotoxicity of ACA-1 on gastric adenocarcinoma cells [AGS] and fibroblasts [HgF] was determined after 24 hours of incubation by MTT assay. Annexin V-FITC and PI staining method was used for measuring the cell apoptosis. Activity of aspase 8 and 9 was assayed by enzymatic method. An ECMatrix was used for determining invasion ability of AGS cells. ACA-1 showed strong and dose - dependent toxicity on AGS cells by induction of early apoptosis. Increase in caspase 8 and 9 activities was involved in this process. Also, ACA-1 decrease the invasion ability on AGS cells. ACA-1 induced apoptosis in human gastric cancer cells by activating caspase 8 and 9. With respect to decreasing cell invasion of AGS cells, ACA-1 may be considered as a potential candidate against human gastric cancer

[Evaluation of coral [madrepora] cytotoxicity in fibroblast cell culture]

Bone defect is one of the major problems facing dentists and maxillofacial surgeons. Several approaches, such as incorporating autografts, xenografts, as well as using polymers have been suggested. The calcified skeleton of corals has been proposed for bone grafting over the past few years. Toxicity is the most important concern of grafting foreign material. The purpose of this study was to investigate the toxicity of coral [madrepora] skeleton in fibroblast culture. Powdered corals with particle sizes of less than 100 micron were autoclaved. Coral powder was added to culture medium consisting of 90% RPMI, 10% FBS and different concentrations of 0.5, 1, 2, 5, 10, 20 and 50 milligrams per 100 milliliters of media were prepared. Fibroblast cells were obtained from Pasteur institute for inserting into culture mediums. Three samples were prepared from each concentration. Complete medium culture was the control. Ninety six well plate containing culture medium, fibroblasts and coral powder were prepared. They were placed into a programmed incubator containing 5% CO2 for 24, 48 and 72 hours at 37°C. The [MTT] was then added to plate culture mediums. Since light absorption has close relationship with cells viability, ELISA reader apparatus was used to measure light absorption [optic dosimeter]. The collected data was statistically analyzed using ANOVA test. Light absorption in control group was similar to light absorption in the various concentrations of coral medium culture along with incorporated fibroblasts. Based on the findings of this study, it can be concluded that coral [madrepora] has potentially no cytotoxic effect on fibroblast cells

Purification of immunomodulator fraction components of garlic [R10] by HPLC

It has been reported that garlic extract and its components show medicinal effects including immunomodulatory activities. We have isolated the immunomodulatory fraction [R10] previously In this study we have proposed to purify the components of R10 using HPLC. Crude garlic extract purchased from Hamadan, Iran. R10 fraction including 10-50 KD molecules have been isolated using ultrafiltration. Further purification has been made using Vaydac 208Tpv10, a semi-preparative HPLC reversphase choromatography column. Tricine SDS-PAGE has been used to determine molecular weight of the samples. 6 major peakes were obtained from HPLC of R10 at gradient of 0.25% of buffer B/min through 60 minute. The molecular weight of 3 peaks [samples 0, 1 and 3] was 12 KD with tricine-SDSPAGE. Using C8 HPLC reversphase choromatography seems to be appropriate tool for purification of R10 components and the purified components at peak 0, 1 and 3 seems to be isotypic molecules

effect of combination of pseudomonas aeruginosa alginate and an immunomodulator protein of garlic on opsonophagocytosis in murine model

Chronic pulmonary infection in patients with cystic fibrosis is predominantly due to infection by mucoid strains of Pseudomonas aeruginosa. Mucoid P. aeriginosa is due to the production of exopolysaccharide called also alginate. Alginate in addition to interference with the clearance of lung has antiphagocytic property. Optimal killing activity of P. aeruginosa requires opsonic antibodies. Since immunomodulatory effects of garlic on enhancing phagocytic activity has been proved, in this study the effect of combination of alginate and an immunomodulator protein of garlic on production of opsonic antibodies against P. aeruginosa mucoid exopolysaccharide has been investigated. Alginate was extracted from a 72-hour culture of P. aeruginosa strain 8821M and then DNasel, RNaseA and Proteinase k were added. Subsequently, alginate was purified with gel filtration chromatography by sephacryl S-400. Female BALB/c mice aged 6-8 weeks were divided into five groups and injected subcutaneously on days 0, 7, 14 with either alginate, garlic, alginate- garlic, R10 or alginate-R10 and opsonophagocytic killing activity was calculated in each group. The purified alginate contained 34.6 micro g/ml uronic acid, 0.5 micro g/ml nucleic acid, 1.45 micro g/ml protein and 0.08 micro g/ml LPS. Opsonophagocytic killing activity after immunization with R10, alginate and their combination showed significant increases of respectively 69%, 67% and 82% comparing to the control group. Combination of P.aeruginosa alginate and immunomodulator fraction of garlic enhances immunogenicity to P.aeroginosa by the elicitation of opsonic antibodies in BALB/c mice

[ cytotoxic effects a of ACA1 on human melanoma cell line]

Different methods such as surgery, chemotherapy, radiotherapy, hormone therapy and immunotherapy are used for treatment of melanoma cancer. Unfortunately they don't always have desirable results and they may have unfavorable side effects. Researchers try to find new, more effective drugs with low side effects. In this study we evaluated the cytotoxic effect of ACA-1, a water extract of a traditional Iranian medicinal herbs on a melanoma cell line SKMEL-3. The SKMEL3 cell line was obtained from Pasture institute, Tehran, Iran and cultured in RPMI media supplemented with 10% FBS. Equal number of cells were added to a 96 well microplate and were incubated with various doses of ACA1 [5,2,1,0.2,0.1,0.05,0.02 and 0.01 mg/ml] for 24, 48 and 72 hours in parallel. The cytotoxic effects of the drug was evaluated using MTT assay. The Results showed that ACA1 has significant cytotoxic effects with dose and time dependent manner on SKMEL3. The optimum dose [5 mg/ml] showed 47% cytotoxicity in 24 h, 65% cytotoxicity in 48 h and 71% cytotoxicity in 72 h. Based on the results of this research, ACA1 is a suitable candidate for chemotherapy of melanoma patients. Further studies are necessary in order to find effective drugs, their effects on other cell lines and approved in vivo models and clinical trials

[Comparison of in vitro cytotoxic effect of two MTAs [Iran made and imported] on peripheral blood mononuclear cells]

Imported mineral Trioxide Aggregate [MTA] as a root -end filling material has many applications in root repairing and bone healing including direct pulp capping, repair of root and furcation perforation, and apexification. By introducing a similar material, made in Iran, namely Root MTA, primary tests such as cytotoxicity test should be performed. The purpose of this study was to compare the 24h and 48h cytotoxicity effect of Root MTA on peripheral blood mononuclear cells using MTT Assay. In this study, after blood sampling, peripheral blood mononuclear cells were isolated on a gradient of ficole hypac. After several washing phases, cells were cultured in RPMI with 10% FCS in 96 well U shape microtiterplates in a 5% Co[2] incubator. Serial dilution of MTA and Root MTA were added to the wells. Cell cytotoxicity was determined, 24 and 48 hours after treatment by MTT Assay. Cytotoxicity of Root MTA was less than that of MTA [ProRoot ]. At 24 hours after culture there was significant difference between cell viability affected by Root MTA and MTA in the three different dilutions 20 lamda, 50 lamda, and 100 lamda [p<0.05]. At 48 hours, there was also a significant difference between cell viability after using Root MTA and MTA in two dilutions 50 lamda, 100 lamda [p<0.05]. Root MTA could be considered as a proper alternative for MTA. For ther studies such as clinical trials should also be performed