ABSTRACT
BACKGROUND: Euphorbia hirta has been reported to possess anti-inflammatory activity. This study was carried out to determine the prostaglandin E 2 (PGE 2 ) inhibition activity of the fractions of the E. hirta aqueous extract on rabbit synovial fibroblast cells (HIG-82). METHODS: E. hirta aqueous extract was fractionated into five fractions (fractions A, B, C, D, and E) by reversed phase flash chromatography. Rabbit synovial fibroblast cells (HIG-82) were activated with phorbol myristate acetate and treated with the fractions. The amount of PGE 2 released into the medium was measured by enzyme-linked immunosorbent assay. RESULTS: Fraction A (0.1, 1, and 10 µg/ml) had the greatest PGE 2 inhibitory effect among the five fractions, and showed a greater extent of PGE 2 inhibition compared to the aqueous extract. In contrast, Fraction E had the greatest stimulatory effect on PGE 2 release. CONCLUSIONS: Fraction A of the aqueous extract inhibited the production of PGE 2 from activated HIG-82 cells to a greater extent than the crude aqueous extract. Bioactive compounds with anti-inflammatory activity are likely to be concentrated in Fraction A of E. hirta aqueous extract.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Dinoprostone/metabolism , Euphorbia/metabolism , Fibroblasts/drug effects , Plant Extracts/pharmacology , Animals , RabbitsABSTRACT
Bacillus thuringiensis (Bt) parasporal proteins with selective anticancer activity have recently garnered interest. This study determines the efficacy and mode of cell death of Bt 18 parasporal proteins against 3 leukemic cell lines (CEM-SS, CCRF-SB and CCRF-HSB-2).Cell-based biochemical analysis aimed to determine cell viability and the percentage of apoptotic cell death in treated cell lines; ultrastructural analysis to study apoptotic changes and Western blot to identify the parasporal proteins' binding site were performed. Bt 18 parasporal proteins moderately decreased viability of leukemic cells but not that of normal human T lymphocytes. Further purification of the proteins showed changes in inhibition selectivity. Phosphatidylserine externalization, active caspase-3, cell cycle, and ultrastructural analysis confirmed apoptotic activity and S-phase cell-cycle arrest. Western blot analysis demonstrated glyceraldehyde 3-phosphate dehydrogenase as a binding protein. We suggest that Bt 18 parasporal proteins inhibit leukemic cell viability by cell-cycle arrest and apoptosis and that glyceraldehyde 3-phosphate dehydrogenase binding initiates apoptosis.
Subject(s)
Apoptosis/drug effects , Bacillus thuringiensis , Bacterial Proteins/pharmacology , Caspase 3/metabolism , Caspase 7/metabolism , Cell Cycle Checkpoints/drug effects , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , B-Lymphocytes/drug effects , B-Lymphocytes/pathology , B-Lymphocytes/ultrastructure , Cell Cycle/drug effects , Cell Line , Endotoxins/pharmacology , Humans , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Precursor Cell Lymphoblastic Leukemia-Lymphoma/metabolism , T-Lymphocytes/drug effects , T-Lymphocytes/pathology , T-Lymphocytes/ultrastructureABSTRACT
This study aimed to assess the inhibitory activities of methanol extracts from the microalgae Ankistrodesmus convolutus, Synechococcus elongatus, and Spirulina platensis against Epstein-Barr virus (EBV) in three Burkitt's lymphoma (BL) cell lines, namely Akata, B95-8, and P3HR-1. The antiviral activity was assessed by quantifying the cell-free EBV DNA using real-time polymerase chain reaction (PCR) technique. The methanol extracts from Ankistrodesmus convolutus and Synechococcus elongatus displayed low cytotoxicity and potent effect in reducing cell-free EBV DNA (EC(50)<0.01 µg/ml) with a high therapeutic index (>28000). After fractionation by column chromatography, the fraction from Synechococcus elongatus (SEF1) reduced the cell-free EBV DNA most effectively (EC(50)=2.9 µg/ml, therapeutic index>69). Upon further fractionation by high performance liquid chromatography (HPLC), the sub-fraction SEF1'a was most active in reducing the cell-free EBV DNA (EC(50)=1.38 µg/ml, therapeutic index>14.5). This study suggests that microalgae could be a potential source of antiviral compounds that can be used against EBV.
Subject(s)
Antiviral Agents/isolation & purification , Antiviral Agents/pharmacology , Herpesvirus 4, Human/drug effects , Microalgae/chemistry , Acyclovir/pharmacology , Base Sequence , Burkitt Lymphoma/drug therapy , Burkitt Lymphoma/virology , Cell Line, Tumor , Chlorophyta/chemistry , DNA, Viral/drug effects , DNA, Viral/genetics , Foscarnet/pharmacology , Herpesvirus 4, Human/genetics , Humans , Lymphoid Progenitor Cells/drug effects , Lymphoid Progenitor Cells/virology , Polymerase Chain Reaction , Spirulina/chemistry , Synechococcus/chemistry , Viral Load/drug effectsABSTRACT
During a study on the quality of the indoor environment, Acanthamoeba spp. were detected in 20 out of 87 dust samples collected from air-conditioners installed in a four-story campus building located in Kuala Lumpur, Malaysia. Twenty-one cloned Acanthamoeba isolates designated as IMU1 to IMU21 were established from the positive primary cultures. Five species were identified from the 16 isolates according to the morphological criteria of Pussard and Pons; i.e. A. castellanii, A. culbertsoni, A. griffini, A. hatchetti and A. polyphaga. Species identities for the remaining five isolates (IMU4, IMU5, IMU15, IMU20 and IMU21), however, could not be determined morphologically. At genotypic characterization, these isolates were placed into T3 (IMU14); T5 (IMU16 and IMU17) and T4 (all the remaining isolates). To predict the potential pathogenicity of these Acanthamoeba isolates, thermo- and osmotolerance tests were employed; many isolates were predicted as potential human pathogens based on the outcome of these tests. This is the first time potentially pathogenic Acanthamoeba have been isolated from air-conditioners in Malaysia.
