A Study of Synergy of Combination of Eosin B with Chloroquine, Artemisinin, and Sulphadoxine-Pyrimethamine on Plasmodium falciparum In Vitro and Plasmodium berghei In Vivo.

METHODS: Drug assessment was carried out singly or in combination on Plasmodium falciparum in vitro using the candle jar method at three inhibitory concentrations. Percent parasitemia of live cells was obtained by microscopic counting. Peter's suppression test was carried out on mice infected with Plasmodium berghei after 3 administration of the drugs singly and in combination, and parasites were counted by microscopy for 10 days. RESULTS: Synergy was exhibited by isobolograms of eosin B combined with artesunate and sulphadoxine-pyrimethamine with more than 10 fold reduction of all drugs in vitro. A good combination index was obtained with artesunate at 50% inibitory concentration with 3.4 nM eosin B and 1.7 nM artesunate in contrast to 124 nM eosin B and 7.6 nM artesunate singly. In vivo studies also showed a considerable lowering of the effective dose of eosin B 30 mg/kg: artesunate 3 mg/kg with 200 mg/kg eosin B and 60 mg/kg artesunate separately. Sulphadoxine-pyrimethamine seemed to have the greatest synergistic effect with a combination index of 0.007, but this could be due to it consisting of a combination of three drugs. Eosin B's combination index with chloroquine was fair, and in vivo tests too did not show as much competence as the other two drugs. Conclusion and Interpretation. It can be concluded that eosin B can be used in combination with antimalarial drugs with favorable results.

An Integrative In Silico Mathematical Modelling Study of The Anti-Cancer Effect of Clove Extract ( Syzygium aromaticum ) Combined with In Vitro Metabolomics Study Using 1HNMR Spectroscopy.

BACKGROUND: Clove oil is known for its medicinal properties. The mechanism of anti-cancer properties of Syzygium aromaticum were investigated by mathematical modelling on the genome scale with metabolomics using 1H Nuclear Magnetic Resonance spectroscopy on Raji cells. OBJECTIVES: An integrative analysis correlated the metabolites identified by 1HNMR and genes with the detected pathways. MATERIALS AND METHODS: Raji cells treated with clove oil were collected and sent for 1HNMR spectroscopy and the spectra analyzed by MATLAB and Human Metabolome Database for metabolite identification. Pathway and topology analysis was implemented using the genes and metabolites in the integrative analysis of Metaboanalyst software. RESULTS: 50% inhibitory concentration of clove oil was 50 µg/ml and the model anticipated 74 genes with differentiating metabolites being some amino acids, cholesterol and fucose. CONCLUSION: The integrative study predicted that the anti cancer mechanism of clove oil involves novel enzymes, as likely drug targets, 24-dehydrocholesterol reductase and 7-dehydrocholesterol reductase in cholesterol biosynthesis, dehydrofolate reductase in one carbon metabolism and serine palmitoyl-transferase long chain in sphingolipid biosynthesis.

Characterization of Glycoproteins of Native 19kDa C-Terminal Merozoite Surface Protein-1 from Native Antigen of Plasmodium falciparum.

BACKGROUND: Plasmodium falciparum is the protozoan parasite which causes malignant malaria of medical concern. Prime candidates for recombinant vaccine development are asexual stage antigens of P. falciparum, for example, merozoite surface proteins (MSP1 and MSP2) not given satisfactory results to date. In this study, the 19kDa C-terminal of MSP1, a vaccine candidate was purified in its native form in the ring stage, and its glycoproteins studied. METHODS: The study was carried out at the Biochemistry Department of Pasteur Institute of Iran in the years 2015-2016. Large scale culture of P. falciparum was performed in vitro with 80% ring stage parasitemia. Isopycnic ultracentrifugation with 36% sucrose and analytical SDS-PAGE on the supernatant and precipitate performed, and the 19kDa antigen was obtained by cutting it from strips of preparative SDS gels. Purified protein was concentrated and analyzed by SDS-PAGE and immunoblotting, using antibodies raised to recombinant C-terminal MSP1. RESULTS: The purified protein gave a single band of 19kDa antigen as shown by silver staining of SDS-PAGE and a single bond in immunoblotting. Bioinformatics also confirmed the likelihood of the presence of glycans on the antigen. CONCLUSION: The presence of N and O-glycoproteins were detected by Q proteome kit. This work was done on the ring stage, and earlier workers confirmed the presence of glycoproteins on MSP1 in the other stages. This glycosylation is present in all stages, and maybe incomplete protection elicited by recombinant MSP1 antigens is due to lack of N and O-glycoproteins.

Staging of colorectal cancer using serum metabolomics with 1HNMR Spectroscopy.

OBJECTIVES: Determination of stages of colon cancer is done by biopsy usually after surgery. Metabolomics is the study of all the metabolites using LC-MS and 1HNMR spectroscopy with chemometric techniques. The stages of colon cancer were detected from patients' sera using 1HNMR. MATERIALS AND METHODS: Five ml blood was collected from 16 confirmed patients referred for colonoscopy. One group of eight patients were diagnosed with stage 0 to I colon cancer and the second group of 8 patients with II-IV stage colon cancer. Sera were sent for 1HNMR. The differentiating metabolites were identified using HMDB and the metabolic cycles from Metaboanalyst. RESULTS: Six metabolites of which pyridoxine levels lowered, and glycine, cholesterol, taurocholic acid, cholesteryl ester and deoxyinosine increased. CONCLUSION: The different stages of cancer were identified by the main metabolic cycles such as primary bile acid biosynthesis, purine and vitamin B metabolic pathways and the glutathione cycle.

Corrigendum to "The Effect of Aqueous Extract of Cinnamon on the Metabolome of Plasmodium falciparum Using (1)HNMR Spectroscopy".

[This corrects the article DOI: 10.1155/2016/3174841.].

The Effect of Aqueous Extract of Cinnamon on the Metabolome of Plasmodium falciparum Using (1)HNMR Spectroscopy.

Malaria is responsible for estimated 584,000 deaths in 2013. Researchers are working on new drugs and medicinal herbs due to drug resistance that is a major problem facing them; the search is on for new medicinal herbs. Cinnamon is the bark of a tree with reported antiparasitic effects. Metabonomics is the simultaneous study of all the metabolites in biological fluids, cells, and tissues detected by high throughput technology. It was decided to determine the mechanism of the effect of aqueous extract of cinnamon on the metabolome of Plasmodium falciparum in vitro using (1)HNMR spectroscopy. Prepared aqueous extract of cinnamon was added to a culture of Plasmodium falciparum 3D7 and its 50% inhibitory concentration determined, and, after collection, their metabolites were extracted and (1)HNMR spectroscopy by NOESY method was done. The spectra were analyzed by chemometric methods. The differentiating metabolites were identified using Human Metabolome Database and the metabolic cycles identified by Metaboanalyst. 50% inhibitory concentration of cinnamon on Plasmodium falciparum was 1.25 mg/mL with p < 0.001. The metabolites were identified as succinic acid, glutathione, L-aspartic acid, beta-alanine, and 2-methylbutyryl glycine. The main metabolic cycles detected were alanine and aspartame and glutamate pathway and pantothenate and coenzyme A biosynthesis and lysine biosynthesis and glutathione metabolism, which are all important as drug targets.

Serum Metabolomic Profiling of Sulphur Mustard-Exposed Individuals Using (1)H Nuclear Magnetic Resonance Spectroscopy.

Sulphur mustard is an alkylating agent that reacts with different cellular components, causing acute and delayed complications that may remain for decades after exposure. This study aimed to identify differentially expressed metabolites between mustard-exposed individuals suffering from chronic complications compared with unexposed individuals as the control group. Serum samples were obtained from 15 mustard-exposed individuals and 15 apparently healthy unexposed individuals. Metabolomic profiling was performed using (1)H nuclear magnetic resonance spectroscopy, and analyses were carried out using Chenomex and MATLAB softwares. Metabolites were identified using Human Metabolome Database, and the main metabolic pathways were identified using MetaboAnalyst software. Chemometric analysis of serum samples identified 11 differentially expressed metabolites between mustard-exposed and unexposed groups. The main pathways that were influenced by sulphur mustard exposure were related to vitamin B6 (down-regulation), bile acid (up-regulation) and tryptophan (down-regulation) metabolism. Metabolism of vitamin B6, bile acids and tryptophan are the most severely impaired pathways in individuals suffering from chronic mustard-induced complications. These findings may find implications in the monitoring of exposed patients and identification of new therapeutic approaches.

A metabolic study on colon cancer using (1)h nuclear magnetic resonance spectroscopy.

Background. Colorectal carcinoma is the third cause of cancer deaths in the world. For diagnosis, invasive methods like colonoscopy and sigmoidoscopy are used, and noninvasive screening tests are not very accurate. We decided to study the potential of (1)HNMR spectroscopy with metabolomics and chemometrics as a preliminary noninvasive test. We obtained a distinguishing pattern of metabolites and metabolic pathways between colon cancer patient and normal. Methods. Sera were obtained from confirmed colon cancer patients and the same number of healthy controls. Samples were sent for (1)HNMR spectroscopy and analysis was carried out Chenomex and MATLAB software. Metabolites were identified using Human Metabolic Data Base (HDMB) and the main metabolic cycles were identified using Metaboanalyst software. Results. 15 metabolites were identified such as pyridoxine, orotidine, and taurocholic acid. Main metabolic cycles involved were the bile acid biosynthesis, vitamin B6 metabolism, methane metabolism, and glutathione metabolism. Discussion. The main detected metabolic cycles were also reported earlier in different cancers. Our observations corroborated earlier studies that suggest the importance of lowering serum LCA/DCA and increasing vitamin B6 intake to help prevent colon cancer. This work can be looked upon as a preliminary step in using (1)HNMR analysis as a screening test before invasive procedures.

Efficacy of eosin B as a new antimalarial drug in a murine model.

The initial success of any adopted anti-infective strategy to malaria is followed by a descent due to the emergence of resistance to it. The search for new drugs and drug targets is a consistent demand in this disease. Eosin B, a common laboratory dye, is reported to have good antiparasitic properties in vitro. It was studied for its antiparasitic effect in vivo on chloroquine-sensitive Plasmodium berghei murine malaria. Eosin B was administered in 2 different doses by either the oral or parenteral route, once or twice daily to mice infected with Plasmodium berghei. Both the doses of eosin B 400 mg/kg and 800 mg/kg gave better results than the controls which were 40 mg/kg chloroquine and 100 mg/kg of arteether with P < 0.005 significance. Percentage suppressive activity by Peter's test of eosin B was better, though at a higher dose than both the controls. Survival rate of mice receiving the higher dose of eosin B was longer than that of the controls. When administered twice daily, the mice were fully cured after 4 days. Eosin B seems to be a promising drug exhibiting good antimalarial effects in the murine model of the disease.