ABSTRACT
Lead is a heavy metal of considerable environmental and occupational concern and there is growing evidence that it is toxic to the human immune system. In this regard, this study examined the effect of lead (Pb) exposure to peritoneal macrophages (Mvarphis) of mice (Mus musculus) cultivated in DMEM medium supplemented with fetal bovine serum, in order to investigate cell damage related to cell death. Cells were exposed to two concentrations of inorganic lead [Pb(II)] for 4, 24 and 72h. Cell viability declined during the treatment, with responses including cell death, cellular damage and DNA damage. Cell death images were found in treated cells with an increase in Bax expression, but the inorganic lead failed to induce the loss of membrane asymmetry (Annexin V conjugates), suggesting that cell death was mainly due to necrosis induction. The effects of Pb(II) on the mechanisms of cell death is not completely understood, but the immunosuppression due to DNA damage and Mvarphis death is discussed here. We have previously shown the effect of inorganic lead in mitochondria and phagocytosis in Mvarphis, suggesting here a pathway for the effect of the metal on mechanisms of cell death, also discussing its effects on the immune system.
Subject(s)
Cell Death/drug effects , DNA Damage , Lead/pharmacology , Macrophages, Peritoneal/cytology , Animals , Cell Survival/drug effects , Cells, Cultured , Comet Assay , Cytoplasmic Vesicles/drug effects , Male , Mice , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Models, Biological , bcl-2-Associated X Protein/biosynthesisABSTRACT
Cell cultures of Mandevilla velutina have proved to be an interesting production system for biomass and secondary metabolites able to inhibit the hypotensive activity of bradykinin, a nonapeptide generated in plasma during tissue trauma. The crude ethyl acetate extract of cultured cells contains about 31- to 79-fold more potent anti-bradykinin compounds (e.g., velutinol A) than that obtained with equivalent extracts of tubers. Somaclonal variation may be an explanation for the wide range of inhibitor activity found in the cell cultures. The heterogeneity concerning morphology, differentiation, carbon dissimilation, and velutinol A production in M. velutina cell cultures is reported. Cell cultures showed an asynchronous growth and cells in distinct developmental stages. Meristematic cells were found as the major type, with several morphological variations. Cell aggregates consisting only of meristematic cells, differentiated cells containing specialized cell structures such as functional chloroplasts (cytodifferentiation) and cells with embryogenetic characteristics were observed. The time course for sucrose metabolism indicated cell populations with significant differences in growth and metabolic rates, with the highest biomass-producing cell line showing a cell cycle 60% shorter and a metabolic rate 33.6% higher than the control (F2 cell population). MALDI-TOF mass spectrometric analysis of velutinol A in selected cell lines demonstrated the existence of velutinol A producing and nonproducing somaclones. These results point to a high genetic heterogeneity in general and also in terms of secondary metabolite content.
Subject(s)
Genetic Variation/genetics , Plant Extracts/chemistry , Plants, Medicinal/genetics , Bradykinin/antagonists & inhibitors , Brazil , Cell Culture Techniques/methods , Cell Line , Chromatography , Meristem/cytology , Microscopy, Electron, Scanning , Phenotype , Plant Extracts/metabolism , Plants, Medicinal/cytology , Plants, Medicinal/metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Sucrose/metabolismABSTRACT
Cell cultures of Mandevilla velutina have proved to be an interesting production system for biomass and secondary metabolites able to inhibit the hypotensive activity of bradykinin, a nonapeptide generated in plasma during tissue trauma. The crude ethyl acetate extract of cultured cells contains about 31- to 79-fold more potent anti-bradykinin compounds (e.g., velutinol A) than that obtained with equivalent extracts of tubers. Somaclonal variation may be an explanation for the wide range of inhibitor activity found in the cell cultures. The heterogeneity concerning morphology, differentiation, carbon dissimilation, and velutinol A production in M. velutina cell cultures is reported. Cell cultures showed an asynchronous growth and cells in distinct developmental stages. Meristematic cells were found as the major type, with several morphological variations. Cell aggregates consisting only of meristematic cells, differentiated cells containing specialized cell structures such as functional chloroplasts (cytodifferentiation) and cells with embryogenetic characteristics were observed. The time course for sucrose metabolism indicated cell populations with significant differences in growth and metabolic rates, with the highest biomass-producing cell line showing a cell cycle 60 percent shorter and a metabolic rate 33.6 percent higher than the control (F2 cell population). MALDI-TOF mass spectrometric analysis of velutinol A in selected cell lines demonstrated the existence of velutinol A producing and nonproducing somaclones. These results point to a high genetic heterogeneity in general and also in terms of secondary metabolite content
Subject(s)
Genetic Variation , Plant Extracts , Plants, Medicinal , Brazil , Cell Culture Techniques , Cell Line , Chromatography , Meristem , Microscopy, Electron, Scanning , Phenotype , Plant Extracts , Plants, Medicinal , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , SucroseABSTRACT
The most active polysaccharides which show anti-tumoral activity are (1-->3)-beta-D-glucans, branched or not at O-6. Since these structures are sometimes poorly soluble in aqueous media, alpha-D-glucans and their chemical derivatives, which are more soluble, were also studied. The present object is to observe morphological alterations in HeLa cells caused by two different polysaccharides obtained from the lichen Ramalina celastri, which are (1-->3),(1-->4)-linked alpha-D-glucan and its sulphated derivative. The cells were incubated in Eagle's medium in the absence or presence of each polysaccharide and routinely processed and analysed by light and electron microscopy. Even though the alpha-D-glucan altered the cellular volume, cytoplasmic densities, and mitosis, the resulting monolayer was similar to the control. TEM analysis showed cytoplasmic blebbing and the presence of an amorphous electron-dense material free in the cytoplasm and interior membranes. The enhanced injury caused by the sulphated derivative was apparent, altering cell adhesion and causing cell aggregation. Nuclear modifications such as fragmentation and condensation of chromatin under the nuclear envelope, which showed to be convoluted, suggested the occurrence of cell death by apoptosis.
Subject(s)
HeLa Cells/drug effects , HeLa Cells/pathology , Lichens/metabolism , Polysaccharides/toxicity , HeLa Cells/ultrastructure , Humans , Microscopy, ElectronABSTRACT
1. Lectins labeled with colloidal gold particles were used for the ultrastructural evaluation of the biological effects of GaAs softlaser irradiation on the healing of dog tendon wounds. 2. Six dogs were submitted to tenotomy and tenorrhaphy on the right and left hind legs. All animals received laser irradiation (4 J/cm2) daily for ten days only on the left leg, and the right leg of the same animal was used as control. Biopsies were taken 11, 22 and 40 days after surgery. 3. Laser-irradiated and control tendon tissues were embedded in L.R. White resin and thin sections were incubated in the presence of gold-labeled Arachis hypogaea (PNA), Canavalia ensiformes (Con A) and Triticum vulgare (WGA). 4. In general, the control and laser-irradiated tissues presented homogeneous and similar labelling of heterochromatin, rough endoplasmic reticulum and extracellular matrix. 5. We conclude that GaAs softlaser irradiation does not produce significant changes in the glycosylation of healing tendons.
Subject(s)
Lasers , Lectins , Tendons/radiation effects , Animals , Collagen/radiation effects , Collagen/ultrastructure , Dogs , Extracellular Matrix/radiation effects , Extracellular Matrix/ultrastructure , Female , Fibroblasts/radiation effects , Fibroblasts/ultrastructure , Male , Microscopy, Electron , Tendons/surgery , Tendons/ultrastructure , Time Factors , Wound Healing/radiation effectsABSTRACT
1. Lectins labeled with colloidal gold particles were used for the ultrastructural evaluation of the biological effects of GaAs softlaserirradiation on the healing of dog tendon wounds. 2. Six dogs were submitted to tenotomy and tenorrhaphy on the right and left hind legs. All animals received laser irradiation (4J/cm²) daily for ten days only on the left leg, and the right leg of the same animal was used as control. Biopsies were taken 11, 22 and 40 days after surgery. 3. Laser-irradiated and control tendon tissues were embedded in L.R. White resin and thin section were incubated in the presence of gold-labeled Arachis hypogaea (PNA), Canavalia ensiformes (Con A) and Triticum vulgare (WGA). 4. In general, the control and laser-irradiated tissues presented homogeneous and similar labelling of heterochromatin, rough endoplasmic reticulum and extracellular matrix. 5. We conclude that GaAs softlaser irradiation does not produce significant changes in the glycosylation of healing tendons
