A Metabolomic Investigation of the Effect of Eosin B on Game-tocyte of Plasmodium falciparum Using 1HNMR Spectroscopy.

BACKGROUND: Recently eosin B was shown to have an effect on the asexual stage of Plasmodium falciparum and in this study, its activity against gametocytes and changes in the culture medium metabolites were investigated using an1HNMR-based metabolomics approach. METHODS: In the Biochemistry Department of Pasteur Institute of Iran in 2017, parasites were cultured and gametocytogenesis induced by heparin and 5% hematocrit. Sexual stage parasites were tested by eosin B in 90 well plates and IC50 determined using Lactate Dehydrogenase assay. Gametocytes were treated by IC50 dose of eosin B and the medium collected in the two groups: with eosin B and controls and sent for 1HNMR spectroscopy. The spectra were analyzed on MATLAB interface and the altered metabolites in the culture medium and eosin-affected biochemical pathways were identified by Human Metabolome Database and Metabo-analyst website. RESULTS: The results revealed eosin B had an effective gametocytocidal activity against P. falciparum. The significant metabolites changed in the medium were thia-mine, Asp, Asn, Tyr, Lys, Ala, Phenylpyruvic acid, NAD+ and lipids. The main pathways identified were aminoacyl-tRNA biosynthesis, Phenylalanine, tyrosine and tryptophan biosynthesis, Alanine, aspartate and glutamate metabolism, Phenylala-nine metabolism, Nicotinate and nicotinamide metabolism, and lysine degradation. CONCLUSION: Eosin B exhibited substantial gametocytocidal activity and affected important drug targets in the Plasmodium.

Inhibitory effects of taraxasterol and aqueous extract of Taraxacum officinale on calcium oxalate crystallization: in vitro study.

OBJECTIVE: We investigated and compared the effects of taraxasterol, aqueous extract of T. officinale (AET) aerial part, and potassium citrate (PC) on calcium oxalate (CaOx) crystallization in vitro. MATERIALS AND METHODS: CaOx crystallization was induced by adding sodium oxalate to synthetic urine. Taraxasterol (2.5, 5, 7.5 and 12.5 µg/mL), extract (1, 2, 4 and 8 mg/mL), and PC (100, 150, 200 and 350 mg/mL) were subjected to anti-crystallization activities. The absorbance and %inhibition of nucleation of CaOx crystals were evaluated by spectrophotometer at 2, 4, 6, 8, 10, 20, 30, 40, 50 and 60 min and the number and morphology of crystals were studied by light microscopy after 60 min. RESULTS: Presence of taraxasterol, extract and PC decreased absorbance in experimental samples compared to control, significantly. The nucleation of crystals is inhibited by taraxasterol, extract, and PC (26-64, 55-63 and 60-70%, respectively). The number of CaOx crystals were decreased in presence of taraxasterol (p < .01), extract (p < .001), and PC (p < .001) in a dose-dependent manner. Presence of taraxasterol, extract, and PC decreased the number of CaC2O4 monohydrate, while increased CaC2O4 dihydrate crystals, significantly. Also, the diameter of CaC2O4 dihydrate crystals was decreased in presence of taraxasterol, extract and PC, significantly. CONCLUSIONS: This research indicated that taraxasterol and extract have anti-crystallization activities and effectiveness of the extract is more potent than taraxasterol. It could be because of another constituent in the extract with the synergistic effect.

Antiurolithiatic effect of the taraxasterol on ethylene glycol induced kidney calculi in male rats.

Taraxasterol is one of the important constituents of Taraxacum officinale L. (Compositae) with antioxidant potential. The present study was designed to evaluate and compare the antiurolithiatic effects of taraxasterol and potassium citrate in the ethylene glycol induced urolithiatic rat. Urolithiasis was induced by ammonium chloride and ethylene glycol in adult male rats. Taraxasterol (2, 4 and 8 mg/kg) and potassium citrate (2.5 g/kg) were treated for 33 days by gavage. Then, the animals were anesthetized and weighted and blood, urine, liver and kidney sampling were done. The kidney sections were prepared by hematoxylin & eosin staining. The liver and kidney coefficients, urine pH, calcium, magnesium, oxalate and citrate levels, serum albumin, calcium and magnesium levels, serum alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase activities, superoxide dismutase and glutathione peroxidase activities in serum, kidney and liver, number of calcium oxalate crystal deposits, score of crystal deposits, score of histopathological damages and score of inflammation in kidney sections were evaluated. The results showed that taraxasterol decreased liver and kidney coefficients (p < 0.001), serum calcium (p < 0.01) level, serum alanine aminotransferase (p < 0.001), aspartate aminotransferase (p < 0.001), lactate dehydrogenase (p < 0.05) activities, urine magnesium (p < 0.05) and oxalate (p < 0.001) levels, number of crystal deposits (p < 0.001), score of crystal deposits (p < 0.01), score of histopathological damages (p < 0.001) and score of inflammation (p < 0.01) in kidney sections, while increased urine pH (p < 0.01), calcium (p < 0.001) and citrate (p < 0.05), serum magnesium (p < 0.001) and albumin (p < 0.01) levels, superoxide dismutase and glutathione peroxidase in serum (p < 0.01), kidney (p < 0.05 and p < 0.001, respectively) and liver (p < 0.01 and p < 0.001, respectively) tissue homogenates in treated urolithiatic rats in comparison to the control urolithiatic rats. The effect of potassium citrate is the same as taraxasterol in treated urolithiatic rats. In conclusion, the effect of taraxasterol could be by improving liver function, changing serum and urine parameters, maintaining the antioxidant environment, reducing crystal deposition, excretion of small deposits from kidney and reducing the chance of them being retained in the urinary tract.

In-silico Metabolome Target Analysis Towards PanC-based Antimycobacterial Agent Discovery.

Mycobacterium tuberculosis, the main cause of tuberculosis (TB), has still remained a global health crisis especially in developing countries. Tuberculosis treatment is a laborious and lengthy process with high risk of noncompliance, cytotoxicity adverse events and drug resistance in patient. Recently, there has been an alarming rise of drug resistant in TB. In this regard, it is an unmet need to develop novel antitubercular medicines that target new or more effective biochemical pathways to prevent drug resistant Mycobacterium. Integrated study of metabolic pathways through in-silico approach played a key role in antimycobacterial design process in this study. Our results suggest that pantothenate synthetase (PanC), anthranilate phosphoribosyl transferase (TrpD) and 3-isopropylmalate dehydratase (LeuD) might be appropriate drug targets. In the next step, in-silico ligand analysis was used for more detailed study of chemical tractability of targets. This was helpful to identify pantothenate synthetase (PanC, Rv3602c) as the best target for antimycobacterial design procedure. Virtual library screening on the best ligand of PanC was then performed for inhibitory ligand design. At the end, five chemical intermediates showed significant inhibition of Mycobacterium bovis with good selectivity indices (SI) ≥10 according to Tuberculosis Antimicrobial Acquisition & Coordinating Facility of US criteria for antimycobacterial screening programs.

Methyl jasmonate influence on silymarin production and plant stress responses in Silybum marianum hairy root cultures in a bioreactor.

In this article our aim was to evaluate mass cultivation of S. marianum hairy roots in a bioreactor to produce silymarin. The effects of methyl jasmonate (MJ) elicitation on the accumulation of silymarin and the extent of the MJ-induced oxidative damage were investigated in bioreactor hairy root cultures of S. marianum. The growth rate of the bioreactor hairy root cultures was higher than that of those in a shake flask after 3 weeks. Silymarin accumulation was increased from 0.13 mg g⁻¹ DW in non-treated hairy roots to 0.22 mg g⁻¹ DW in hairy roots 72 h after 100 µM MJ treatment. Guaiacol peroxidase and ascorbate peroxidase were activated by MJ 72 h after treatment, being 3.2- and 1.3-fold higher, respectively, than that of the control. An increase in enzymatic activity suggests increased scavenging of reactive oxygen species, indicating the tolerance to MJ stress. These results suggest that MJ elicitation is beneficial for silymarin production using bioreactor hairy root cultures.

Identification of cyclophilin as a novel allergen from Platanus orientalis pollens by mass spectrometry.

Oriental plane trees are an important source of airborne allergens in cities of southwest Asia. In spite of extensive studies on Platanus acerifolia allergy, there are no reports on the molecular characterization of pollen allergens from Platanus orientalis trees. In this study, a newly recognized member of cyclophilin family with a molecular weight of 18 kDa was identified as being partly responsible for IgE reactivity of P. orientalis pollen extract.

Oriental plane pollen allergy: identification of allergens and cross-reactivity between relevant species.

Pollen from different tree and grass species represent the greatest inducers of type I allergy worldwide. Oriental plane trees, as Platanus orientalis, are an important source of airborne allergens in cities of the southwest Asia and southeast Europe. This study was aimed to identify relevant allergens of P. orientalis pollen and to ascertain whether P. orientalis allergens have cross-reactivity with related plane trees, such as Platanus acerifolia and Platanus occidentalis pollen components. Nineteen patients with a clinical history of reaction to P. orientalis pollen and a positive skin-prick test (SPT) to P. orientalis pollen extract were included in this study. Identification of IgE-binding proteins in Platanus pollen extracts was elucidated by immunoblotting using sera from P. orientalis pollen-sensitive patients. Cross-reactivity studies among P. orientalis allergens and relevant species was evaluated by immunoblot-inhibition and ELISA inhibition assays. All the patients were polysensitive and exhibited positive SPT to grass and tree pollen allergens. The IgE-binding pattern of P. orientalis pollen extract was well defined. The inhibition experiments showed that the capacity of P. occidentalis was greater than P. acerifolia pollen extract in preventing the reactivity of immobilized P. orientalis pollen extract with the IgE antibodies of patients. The 28-kDa molecule was the most frequent allergen among nine IgE-binding proteins of P. orientalis pollen. The IgE-reactive components of P. orientalis pollen showed a higher level of cross-reactivity with P. occidentalis pollen in comparison with P. acerifolia pollen.