ABSTRACT
Doxorubicin (DOX) is an anticancer drug which is used for treatment of several types of cancers. But the clinical use of doxorubicin is limited because of its cardiotoxicity and cardiomyopathy. Mitochondrial-dependent oxidative stress and cardiac inflammation appear to be involved in doxorubicin-induced cardiotoxicity. Betanin as a bioactive compound in Beetroot (Beta vulgaris L.) displays anti-radical, antioxidant gene regulatory and cardioprotective activities. In this current study, we investigated the protective effect of betanin on doxorubicin-induced cytotoxicity and mitochondrial-dependent oxidative stress in isolated cardiomyocytes and mitochondria. Isolated cardiomyocytes and mitochondria were treated with three concentrations of betanin (1, 5 and 10 µM) and doxorubicin (3.5 µM) for 6 h. The parameters of cellular and mitochondrial toxicity were analyzed using biochemical and flow cytometric methods. Our results showed a significant toxicity in isolated cardiomyocytes and mitochondria in presence of doxorubicin which was related to reactive oxygen species (ROS) formation, increase in malondialdehyde (MDA), increase in oxidation of GSH to GSSG, lysosomal/mitochondrial damages and mitochondrial swelling. While betanin pretreatment reverted doxorubicin-induced cytotoxicity and oxidative stress in isolated cardiomyocytes and mitochondria. These results suggest that betanin elicited a typical protective effect on doxorubicin-induced cytotoxicity and oxidative stress. It is possible that betanin could be used as a useful adjuvant in combination with doxorubicin chemotherapy for reduction of cardiotoxicity and cardiomyopathy.
Subject(s)
Antibiotics, Antineoplastic/toxicity , Betacyanins/pharmacology , Doxorubicin/toxicity , Myocytes, Cardiac/drug effects , Animals , Cell Survival/drug effects , Cells, Cultured , Cytoprotection/drug effects , Glutathione/metabolism , Male , Malondialdehyde/metabolism , Membrane Potential, Mitochondrial/drug effects , Mitochondria, Heart/drug effects , Mitochondria, Heart/metabolism , Myocytes, Cardiac/metabolism , Rats, Wistar , Reactive Oxygen Species/metabolismABSTRACT
In a survey in 2005 some symptoms were observed on three olive trees in a collection of different olive cultivars in one of the central provinces in Iran (Yazd Province). Affected trees exhibited a variable range of symptoms: dwarfing, shoot proliferation, internode shortening, small leaves, leaf rolling, abortive buds, appearance of very small shoots, hypertrophied, and die- back. The possibility of association of a phytoplasma with the disease was evaluated using polymerase chain reaction (PCR). Total DNA was extracted from midribs of symptomatic and healthy olive plants using phytoplasma enriched procedure and tested for the presence of phytoplasma by nested-PCR using phytoplasmal- universal primers R16F2/R16R2, followed by primers fU5/rU3 which amplify 1200 and 880bp DNA fragments of 16SrDNA, respectively. PCR resulted in amplification of expected DNA fragments in symptomatic but not in healthy olive by both primer pairs. On the basis of symptoms and positive reaction in PCR, these plants can be infected with a phytoplasma. This is the first report of olive trees phytoplasmal disease in Iran. Identification of the agent and its comparison with other phytoplasmal isolates reported from other plants (hosts) in Iran are being studied. Based on the quarantine requirements, one of these trees was eradicated and two of them were transplanted in an isolated green house for further studies.
Subject(s)
Olea/microbiology , Phytoplasma/isolation & purification , Plant Diseases/microbiology , Botany/methods , DNA Primers , DNA, Fungal/genetics , DNA, Fungal/isolation & purification , Flowers/microbiology , Iran , Phylogeny , Phytoplasma/classification , Phytoplasma/genetics , Plant Leaves/microbiology , Plant Stems/microbiology , SeasonsABSTRACT
Soybean mosaic virus (SMV) is an important disease in soybean and is widely distributed in northern of Iran. SMV transmitted by soybean seed and detection of it is very important for disease management. In this study, several detection methods including DAS-ELISA, indirect-ELISA, tissue-print immunoassay (TPIA) and Dot immunobinding assay (DIBA) were optimized and compared with each other to identify the virus, using polyclonal antibody. For TPIA, nitrocellulose membrane was used to imprint fresh sections of healthy and infected plant materials, and for DIBA 10 microl of extracts was doted onto nitrocellulose membranes. Both membranes were incubated 1 hour in blocking buffer, and then incubated 2 h in 1:1000 dilution of IgG-conjugate. After incubation the membranes were washed three times with PBS-T buffer for 15 min. Then the membranes were incubated in substrate solution containing NBT/BCIP. After some minutes prints or blots of infected tissues turned dark violet, whereas prints or blots of healthy ones did not show any color changes. In some cases, substrate solution was Fast red, containing 0.2M Tris-HCl buffer and 2mM MgCl2, pH = 7.8, producing red color in infected prints or blots. Both methods are simple and TPIA is rapidly and easily applicable in the field. However, TPIA had some advantages over the others. TPIA is time-saving as there is no need for conventional sap extraction and also nitrocellulose membranes used for printing can be used in the field and stored for a long time or transported to another laboratory for process. These two methods can be used routinely for detection of SMV in many samples.
Subject(s)
Glycine max/virology , Mosaic Viruses/pathogenicity , Plant Diseases/microbiology , Enzyme-Linked Immunosorbent Assay , Immunoassay/methods , Mosaic Viruses/classification , Mosaic Viruses/isolation & purification , SerotypingABSTRACT
Chromosomal instability was examined in 20 apparently healthy children of survivors of childhood malignancy. As compared to controls, no increase of spontaneous or bleomycin-induced aberrations (including gaps, breaks, sister chromatid exchanges, pulverization, and premature centromere divisions) were found in these "index children." The results suggest that the offspring of subjects previously receiving chemotherapy and/or radiotherapy for childhood malignancy are probably at no increased risk of latent chromosomal instability.
Subject(s)
Chromosome Aberrations , Genetic Predisposition to Disease , Neoplasms/genetics , Bleomycin/adverse effects , Centromere/genetics , Child , Female , Humans , Karyotyping , Leukemia/genetics , Lymphoma/genetics , Male , Nuclear Family , Sister Chromatid Exchange , SurvivorsABSTRACT
Anthropometric, dysmorphologic, and cytogenetic investigations of 21 children of 20 survivors of childhood malignancy revealed no signs of congenital anomalies in any of the subjects examined. No increase of mild errors of morphogenesis (minor anomalies) was observed in the well-developing children; no latent chromosome instability was found in their Bleomycin-treated lymphocyte cultures either. The suggestion that previous oncological therapy does not lead to an increased risk of congenital disorders in the offspring was confirmed by the present findings obtained with various, in part new methods.
Subject(s)
Neoplasms/genetics , Anthropometry , Bleomycin/pharmacology , Child , Child, Preschool , Chromosome Aberrations , Cytogenetics , Female , Humans , Infant , Lymphocyte Culture Test, Mixed/methods , Lymphocytes/drug effects , Male , Morphogenesis , Neoplasms/congenital , SurvivorsABSTRACT
The possible effect of in vivo oxygen exposure on chromosomes was examined in lymphocyte cultures of 12 very-low-birthweight infants on the 1st, 8th, and 16th days of intensive care. No increase of cytogenetic anomalies was seen in untreated and bleomycin-treated cultures. The findings suggest that neonatal oxygen exposure is unlikely to cause latent chromosome damage.