ABSTRACT
OBJECTIVES: Apoptosis is the process of programmed cell death (PCD) that occurs in both animal and plant cells. Protozoan parasites possess metacaspase and these caspase-related proteases could be involved in the PCD pathways in these organisms. Therefore we analyzed the activities of metacaspase and PARP genes in Leishmania infantum (MCAN/IR/96/LON49) treated with miltefosine. MATERIALS AND METHODS: Anti-leishmania activity of miltefosine was studied by treatment of cultured promastigotes with various concentration of miltefosine. MTT assay and Annexin-V FLUOS staining by using FACS flow cytometry methods were used. Cytotoxic potential of HePC on the amastigots of L.infantum was evaluated in J774 cell line. In addition, metacaspase and PARP genes expression of treated L. infantum were studied. RESULTS: Miltefosine led to dose-dependent death of L. infantumwith features compatible with apoptosis. Over expression of metacaspase and PARP was seen 6 hr after treatment. CONCLUSIONS: Our study showed that miltefosine exerts cytotoxic effect on L. infantum via an apoptotic-related mechanism.
Subject(s)
Antiprotozoal Agents/pharmacology , Apoptosis/drug effects , Caspases/drug effects , Leishmania infantum/drug effects , Phosphorylcholine/analogs & derivatives , Poly(ADP-ribose) Polymerases/drug effects , Apoptosis/genetics , Colorimetry , Caspases/genetics , Flow Cytometry , Formazans/metabolism , Leishmania infantum/cytology , Leishmania infantum/genetics , Polymerase Chain Reaction , Phosphorylcholine/pharmacology , Poly(ADP-ribose) Polymerases/genetics , Tetrazolium Salts/metabolismABSTRACT
Accurate estimation of the exposure-response relationship between ambient urban particulate matters (PM) and public health is important for regulatory perspective of ambient urban particulate matters (PM). Ambient PM contains various transition metals and organic compounds. PM10 (aerodynamic diameter less than 10 microgram) is known to induce diverse diseases such as chronic cough, bronchitis, chest illness, etc. However, recent evaluation of PM2.5 (aerodynamic diameter less than 2.5 microgram) against health outcomes has suggested that the fine particles may be more closely associated with adverse respiratory health effects than particles of larger size. This study was performed to evaluate PM2.5-induced oxidative stress in rat lung epithelial cell in order to provide basic data for the risk assessment of PM2.5. PM2.5 showed higher cytotoxicity than PM10. Also, PM 2.5 induced more malondialdehyde (MDA) formation than PM10. In Hoechst 33258 dye staining and DNA fragmentation assay, apopotic changes were clearly detected in PM2.5 treated cells in compared to PM10. Expression of catalase mRNA was increased by PM2.5 rather than PM10. PM2.5 induced higher Mth1 mRNA than PM10. In pBR322 DNA treated with PM2.5, production of single strand breakage of DNA was higher than that of PM10. In Western blot analysis, PM2.5 induced more Nrf-2 protein, associated with diverse transcriptional and anti-oxidative stress enzymes, compared to PM10. Our data suggest that PM2.5 rather than PM10 may be responsible for PM-induced toxicity. Additional efforts are needed to establish the environmental standard of PM2.5.
Subject(s)
Animals , Rats , Air Pollutants/chemistry , Apoptosis/physiology , Benzimidazoles/metabolism , Blotting, Western , Cell Line , Cell Survival/physiology , DNA Fragmentation/physiology , DNA Repair Enzymes/genetics , DNA-Binding Proteins/metabolism , Epithelial Cells/drug effects , Formazans/metabolism , GA-Binding Protein Transcription Factor , Lipid Peroxides/metabolism , Lung Diseases/chemically induced , Oxidative Stress/physiology , RNA, Messenger/chemistry , Reverse Transcriptase Polymerase Chain Reaction , Tetrazolium Salts/metabolism , Transcription Factors/metabolismABSTRACT
Taxol is a clinically useful anticancer drug against a variety of cancers. Although it has been known that taxol induces the apoptosis of cancer cells through cytochrome C release and the activation of caspases, the effect of taxol on dendritic cells (DCs) has not been studied. In this study, taxol enhanced the expression of MHC class II on DCs, compared to medium-treated immature DCs. Surprisingly, the viability of DCs was not decreased by taxol, whereas that of cancer cells was. It was confirmed that taxol did not induce the apoptosis of DCs based on annexin V-FITC/propidium iodide (PI) staining assay. Since previous study demonstrated that taxol induced the production of nitric oxide (NO) related to the viability of DCs, the level of NO from taxol-treated DCs was determined. Any significant amount of NO was not detected. Although taxol enhanced the expression of a maturation marker, MHC class II molecules, it strikingly inhibited the proliferation of splenic T lymphocytes activated by DCs. Taken together, this study demonstrated that taxol induced an altered maturation of DCs, the increase of MHC class II molecule but the inhibition of proliferation of splenic T lymphocytes. It is suggested that taxol may induce the immunosuppression in patients with cancer by the inhibition of DC-activated T cell proliferation, but not by the direct killing of DCs.
Subject(s)
Animals , Female , Mice , Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Coloring Agents/metabolism , Dendritic Cells/cytology , Flow Cytometry , Formazans/metabolism , Histocompatibility Antigens Class II/biosynthesis , Lymphocyte Activation/drug effects , Mice, Inbred BALB C , Mice, Inbred C57BL , Nitric Oxide/metabolism , Paclitaxel/pharmacology , T-Lymphocytes/cytology , Tetrazolium Salts/metabolismABSTRACT
Thin layer chromatography of aqueous extract of whole Cheilanthesfarinosa fern indicated the presence of ptaquiloside or ptaquiloside like compound, coinciding Rf values with that of Pterosin B standard. HPLC analysis revealed the presence of 26.3 mg/kg ptaquiloside. In vitro studies of the aqueous extract on lymphocyte culture revealed a correlation between stimulative indices and concentration of aqueous extract. Stimulation in lymphocyte proliferation was in order of bracken > cheilanthes > ConA> ptaquiloside standard. On incubation of lymphocyte with aqueous extract of ferns, no DNA damage was observed in isolated DNA.
Subject(s)
Animals , Cattle , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , DNA Fragmentation/drug effects , Ferns/chemistry , Formazans/metabolism , Lymphocyte Activation/drug effects , Lymphocytes/drug effects , Plant Extracts/toxicity , Plants, Toxic/chemistry , Tetrazolium Salts/metabolismABSTRACT
In the present study, a rapid and simple colorimetric technique has been described to determine the presence of bacteria in tissue culture medium used in animal cell culture. The microplate assay is based on utilization of MTT [3(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide] by bacteria resulting in formation of formazan crystals which can be measured colorimetrically. Contaminated medium, a standard gram-negative and gram-positive bacteria produce formazan from MTT which is related to the bacterial load. The assay has utility in screening tissue culture reagents to detect the presence of bacteria.
Subject(s)
Bacteria/isolation & purification , Bacteriological Techniques/methods , Crystallization , Culture Media/analysis , Drug Contamination , Formazans/metabolism , Tetrazolium Salts/metabolism , Thiazoles/metabolismABSTRACT
Amphetamine, ephedrine, mescaline and chlorpromazine act as substrate for dehydrogenase system of rat brain homogenate. This system appears quite different from the usual deaminating process of these compounds, which required NADP and oxygen. The present system proceeds more rapidly in anaerobic condition.