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Uptake and detoxification of diesel oil by a tropical soil Actinomycete Gordonia amicalis HS-11: Cellular responses and degradation perspectives
2020
Sowani, Harshada | Kulkarni, Mohan | Zinjarde, Smita
A tropical soil Actinomycete, Gordonia amicalis HS-11, has been previously demonstrated to degrade unsaturated and saturated hydrocarbons (squalene and n-hexadecane, respectively) in an effective manner. In present study, G. amicalis HS-11 degraded 92.85 ± 3.42% of the provided diesel oil [1% (v/v)] after 16 days of aerobic incubation. The effect of different culture conditions such as carbon source, nitrogen source, pH, temperature, and aeration on degradation was studied. During degradation, this Actinomycete synthesized surface active compounds (SACs) in an extracellular manner that brought about a reduction in surface tension from 69 ± 2.1 to 30 ± 1.1 mN m⁻¹ after 16 days. The morphology of cells grown on diesel was monitored by using a Field Emission Scanning Electron Microscope. Diesel-grown cells were longer and clumped with smooth surfaces, possibly due to the secretion of SACs. The interaction between the cells and diesel oil was studied by Confocal Laser Scanning Microscope. Some cells were adherent on small diesel droplets and others were present in the non-attached form thus confirming the emulsification ability of this organism. The fatty acid profiles of the organism grown on diesel oil for 48 h were different from those on Luria Bertani Broth. The genotoxicity and cytotoxicity of diesel oil before and after degradation were determined. Cytogenetic parameters such as mitotic index (MI); mitosis distribution and chromosomal aberration (type and frequency) were assessed. Oxidative stress was evaluated by measuring levels of catalase, superoxide dismutase and concentration of malondialdehyde. On the basis of these studies it was deduced that the degradation metabolites were relatively non-toxic.
Показать больше [+] Меньше [-]Whole-transcriptome analysis of the toxic effects of zearalenone exposure on ceRNA networks in porcine granulosa cells
2020
Zearalenone (ZEA), an estrogen-like mycotoxin, is commonly detected in animal feeds including improperly stored grains. It has been well demonstrated that ovarian granulosa cells (GCs) perform vital roles during follicular development, however, the competing endogenous RNA (ceRNA) network in GCs after ZEA exposure remains to be well described. Here, for the first time, we adopted whole-transcriptome sequence technology to explore the molecular mechanism of ZEA toxicology on porcine GCs. The results provide evidence that the cell cycle of porcine GCs is arrested in the G2/M phase after exposure to ZEA. Furthermore, bioinformation analysis found that cell cycle arrest related genes were perturbed, including CDK1, CCNB1, CDC25A, and CDC25C, which was consistent with the results of RT-qPCR, immunofluorescence, and Western Blotting. Based on the whole-transcriptome sequence data, by constructing ceRNA networks related to cell cycle arrest, we observed that ZEA exposure arrested cell cycle progression at the G2/M phase in porcine GCs, and non-coding RNAs (ncRNAs) played an important role in this process via regulating the expressions of cell cycle arrest related genes. Taken together, our data here provides strong data to support that the toxicological mechanism regarding the widely distributed toxicant ZEA acts through ceRNA networks in porcine granulosa cells.
Показать больше [+] Меньше [-]Visible-light reduced silver nanoparticles’ toxicity in Allium cepa test system
2020
Souza, Irisdoris R. | Silva, Lucas R. | Fernandes, Letícia S.P. | Salgado, Lilian D. | Silva de Assis, Helena C. | Firak, Daniele S. | Bach, Larissa | Santos-Filho, Ronaldo | Voigt, Carmen L. | Barros, Ariana C. | Peralta-Zamora, Patricio | Mattoso, Ney | Franco, Celia Regina C. | Soares Medeiros, Lia C. | Marcon, Bruna H. | Cestari, Marta M. | Sant’Anna-Santos, Bruno F. | Leme, Daniela M.
Silver nanoparticles (AgNPs) are widely used in consumer products due to their antibacterial property; however, their potential toxicity and release into the environment raises concern. Based on the limited understanding of AgNPs aggregation behavior, this study aimed to investigate the toxicity of uncoated (uc-AgNP) and coated with polyvinylpyrrolidone (PVP-AgNP), at low concentrations (0.5–100 ng/mL), under dark and visible-light exposure, using a plant test system. We exposed Allium cepa seeds to both types of AgNPs for 4–5 days to evaluate several toxicity endpoints. AgNPs did not cause acute toxicity (i.e., inhibition of seed germination and root development), but caused genotoxicity and biochemical alterations in oxidative stress parameters (lipid peroxidation) and activities of antioxidant enzymes (superoxide dismutase and catalase) in light and dark conditions. However, the light exposure decreased the rate of chromosomal aberration and micronuclei up to 5.60x in uc-AgNP and 2.01x in PVP-AgNP, and 2.69x in uc-AgNP and 3.70x in PVP-AgNP, respectively. Thus, light exposure reduced the overall genotoxicity of these AgNPs. In addition, mitotic index alterations and morphoanatomical changes in meristematic cells were observed only in the dark condition at the highest concentrations, demonstrating that light also reduces AgNPs cytotoxicity. The light-dependent aggregation of AgNPs may have reduced toxicity by reducing the uptake of these NPs by the cells. Our findings demonstrate that AgNPs can be genotoxic, cytotoxic and induce morphoanatomical and biochemical changes in A. cepa roots even at low concentrations, and that visible-light alters their aggregation state, and decreases their toxicity. We suggest that visible light can be an alternative treatment to remediate AgNP residues, minimizing their toxicity and environmental risks.
Показать больше [+] Меньше [-]Silica nanoparticles induce spermatogenesis disorders via L3MBTL2-DNA damage-p53 apoptosis and RNF8-ubH2A/ubH2B pathway in mice
2020
Liu, Jianhui | Li, Xiangyang | Zhou, Guiqing | Sang, Yujian | Zhang, Yue | Zhao, Yanzhi | Ge, Wei | Sun, Zhiwei | Zhou, Xianqing
Silica nanoparticles (SiNPs) can reduce both quality and quantity of sperm via inhibiting the progress of meiosis and mitosis and inducing apoptosis of spermatogenic cells, however, their specific mechanism and effects on the later stage of spermatogenesis are still unclear. To investigate the effects of SiNPs on the reproductive system, male mice were treated with SiNPs (0, 1.25, 5 and 20 mg/kg.bw) via intratracheal instillation once every 3 days and for a total of 15 days. Results revealed that exposure to SiNPs induced reduction in the rate of sperm activity, histological abnormalities in seminiferous epithelium as well as apoptosis of spermatogenic cells, which are associated with decreased level of Lethal (3) malignant brain tumor like 2 (L3MBTL2) and activation of DNA damage-p53-mitochondrial apoptosis pathways. Moreover, reduction in L3MBTL2 level caused by SiNPs also led to the lower expression of RNF8-ubH2A/ubH2B pathway, thus resulting in incomplete histone-to-protamine exchange. These results suggest that the inhibition of L3MBTL2 expression caused by SiNPs not only activates DNA damage-p53-mitochondrial apoptosis pathway leading to the apoptosis of spermatogenic cells, but also inhibits RNF8-ubH2A/ubH2B pathway resulting in incomplete histone-to-protamine exchange, thereby affected spermatogenesis. This indicates that L3MBTL2 plays an important role in reproductive toxicity of males caused by SiNPs.
Показать больше [+] Меньше [-]Long-term exposure of xenoestrogens with environmental relevant concentrations disrupted spermatogenesis of zebrafish through altering sex hormone balance, stimulating germ cell proliferation, meiosis and enhancing apoptosis
2019
Wang, Ya–Qin | Li, Ying–Wen | Chen, Qi–Liang | Liu, Zhi–Hao
Environmental estrogens are capable of interfering with the spermatogenesis and fertility of fish. However in natural waters, these chemicals are more likely to occur as a combination rather than a single stressor. Whether and how the mixture of xenoestrogens with environmental relevant concentrations may affect fish spermatogenesis remains largely unknown. In this study, male zebrafish adults were administered to 17alpha-ethinylestradiol (EE2) and a mixture of xenoestrogens (Mix (E2, EE2, DES, 4-t-OP, 4-NP and BPA)), with the estrogenic potency equivalent to EE2. After a 60-day exposures, elevated mRNA levels of vitellogenin 1 (vtg1) and estrogen receptor 1 (esr1) in the liver of fish in both treated groups were observed. Moreover, the plasma level of E2 declined significantly in the Mix group and the ratio of 11-KT/E2 was significantly elevated in both treated groups. Consistently, the mRNA level of P450 side-chain cleavage (scc) in the EE2 group and ovarian type aromatase (cyp19a1a) in the Mix group was significantly suppressed. In addition, decreased gonadosomatic index and sperm count in the fish of Mix group were present. Furthermore, increased number of the proliferating germ cells (such as spermatogonia and spermatocytes) was observed in the fish of both groups, suggesting a stimulated germ cell proliferation and meiosis. Accordingly, both exposures significantly up-regulated the mRNA levels of genes in mitosis (cyclinb1) and meiosis (cyp26a1 in EE2 group, aldh1a2, cyp26a1, sycp3 and spo11 in Mix). In addition, decreased number of spermatozoa and increased number of TUNEL-positive signals were present in the testis of fish in the Mix group, indicating an enhanced apoptosis. Further analyses demonstrated the significant elevated expressions of tnfrsf1a and the ratio of tnfrsf1a/tnfrsf1b in the Mix group, suggesting an elevated apoptosis in the testis of fish in the Mix group via extrinsic pathway. The present study greatly extends our understanding of the underlying mechanisms of the reproductive toxicity of xenoestrogens on fish.
Показать больше [+] Меньше [-]Endosulfan inhibits proliferation through the Notch signaling pathway in human umbilical vein endothelial cells
2016
Wei, Jialiu | Zhang, Lianshuang | Ren, Lihua | Zhang, Jin | Yu, Yang | Wang, Ji | Duan, Junchao | Peng, Cheng | Sun, Zhiwei | Zhou, Xianqing
Our previous research showed that endosulfan triggers the extrinsic coagulation pathway by damaging endothelial cells and causes hypercoagulation of blood. To identify the mechanism of endosulfan-impaired endothelial cells, we treated human umbilical vein endothelial cells (HUVECs) with different concentrations of endosulfan, with and without an inhibitor for Notch, N-[N-(3, 5-difluorophenacetyl)-1-alanyl]S-Phenylglycinet-butylester (DAPT, 20 μM), or a reactive oxygen species (ROS) scavenger, N-Acetyl-l-cysteine (NAC, 3 mM), for 24 h. The results showed that endosulfan could inhibit cell viability/proliferation by increasing the release of lactate dehydrogenase (LDH), arresting the cell cycle in both S and G2/M phases, and inducing apoptosis in HUVECs. We also found that endosulfan can damage microfilaments, microtubules, and nuclei; arrest mitosis; remarkably increase the expressions of Dll4, Notch1, Cleaved-Notch1, Jagged1, Notch4, Hes1, and p21; and significantly induce ROS and malondialdehyde production in HUVECs. The presence of DAPT antagonized the above changes of cycle arrest, proliferation inhibition, and expressions of Dll4, Notch1, Cleaved-Notch1, Hes1, and p21 caused by endosulfan; however, NAC could attenuate LDH release; ROS and malondialdehyde production; apoptosis; and the expression levels of Dll4, Notch1, Cleaved-Notch1, Notch4, and Hes1 induced by endosulfan. These results demonstrated that endosulfan inhibited proliferation through the Notch signaling pathway as a result of oxidative stress. In addition, endosulfan can damage the cytoskeleton and block mitosis, which may add another layer of toxic effects on endothelial cells.
Показать больше [+] Меньше [-]The Tradescantia micronucleus assay is a highly sensitive tool for the detection of low levels of radioactivity in environmental samples
2016
Mišík, Miroslav | Krupitza, Georg | Mišíková, Katarína | Mičieta, Karol | Nersesyan, Armen | Kundi, M. (Michael) | Knasmueller, Siegfried
Environmental contamination with radioactive materials of geogenic and anthropogenic origin is a global problem. A variety of mutagenicity test procedures has been developed which enable the detection of DNA damage caused by ionizing radiation which plays a key role in the adverse effects caused by radioisotopes. In the present study, we investigated the usefulness of the Tradescantia micronucleus test (the most widely used plant based genotoxicity bioassay) for the detection of genetic damage caused by environmental samples and a human artifact (ceramic plate) which contained radioactive elements. We compared the results obtained with different exposure protocols and found that direct exposure of the inflorescences is more sensitive and that the number of micronuclei can be further increased under “wet” conditions. The lowest dose rate which caused a significant effect was 1.2 μGy/h (10 h). Comparisons with the results obtained with other systems (i.e. with mitotic cells of higher plants, molluscs, insects, fish and human lymphocytes) show that the Tradescantia MN assay is one to three orders of magnitude more sensitive as other models, which are currently available. Taken together, our findings indicate that this method is due to its high sensitivity a unique tool, which can be used for environmental biomonitoring in radiation polluted areas.
Показать больше [+] Меньше [-]Effects of lead on root growth, cell division, and nucleolus of Allium cepa
1994
Liu, D.H. | Jiang, W.S. | Wang, W. | Zhao, F.M. | Lu, C. (Department of Biology, Tianjin Normal University, Tianjin 300074 (China))
Comparative investigation of toxicity induced by UV-A and UV-C radiation using Allium test
2022
Çavuşoğlu, Kültiğin | Kalefetoğlu Macar, Tuğçe | Macar, Oksal | Çavuşoğlu, Dilek | Yalçın, Emine
Organisms are increasingly exposed to ultraviolet (UV) rays of sunlight, due to the thinning of the ozone layer and its widespread use in sterilization processes, especially against the SARS-CoV-2 virus. The present study was conducted with the purpose of evaluating the damages of UV-A and UV-C radiations in Allium cepa L. roots. The effects of two different types of UV on some physiological, biochemical, cytogenotoxic, and anatomical parameters were investigated in a multifaceted study. Three groups were formed from Allium bulbs, one of which was the control group. One of the other groups was exposed to 254 nm (UV-C) and the other to 365 nm (UV-A) UV. Growth retardation effect of UV was investigated with respect to germination percentage, total weight gain, and root elongation, while cytogenotoxicity arisen from UV exposure was analyzed using mitotic index (MI) and chromosomal aberration (CA) and micronucleus (MN) frequency. Oxidative stress due to UV application was investigated based on the accumulation of malondialdehyde (MDA) and the total activities of superoxide dismutase (SOD) and catalase (CAT) enzymes. Also, anatomical changes induced by UV-A and UV-C were analyzed in root meristematic cells. UV treatments caused significant reductions in growth-related parameters. Both UV treatments caused a significant increase in MDA levels and induction of SOD and CAT enzymes in root meristematic cells. A decrease in MI and an increase in the frequency of MN and CAs were observed in root tip cells, indicating the cytogenotoxic effect of UV application. Anatomical damages such as epidermis cell damage, cortex cell damage, necrotic zones, giant cell nucleus, and indistinct transmission tissue occurred in cells exposed to UV. All of the physiological, biochemical, cytogenetic, and anatomical damages observed in this study were more severe in cells treated with UV-C compared to UV-A. This study suggested that UV exposure triggered growth inhibition, cytogenotoxicity, oxidative stress, and meristematic cell damages in A. cepa roots depending on the wavelength.
Показать больше [+] Меньше [-]Protective role of green tea against paraquat toxicity in Allium cepa L.: physiological, cytogenetic, biochemical, and anatomical assessment
2022
Yirmibeş, Ferhat | Yalçin, Emine | Çavuşoğlu, Kültiğin
In this study, the toxic effects of paraquat, one of the most commercially sold herbicides in the world, and the protective role of green tea leaf extract (GTLE) against these effects were investigated. Allium cepa L. bulbs (n = 16) were used as test material. One hundred milligrams per liter dose of paraquat and 190 and 380 mg/L doses of GTLE were preferred. Paraquat toxicity was investigated with the help of physiological (percent germination, root length, and weight gain), cytogenetic (mitotic index = MI, micronucleus = MN, and chromosomal damages = CAs), biochemical (superoxide dismutase = SOD, catalase = CAT, malondialdehyde = MDA), and anatomical (meristematic cell damages) parameters. A. cepa bulbs were divided into 6 groups as 1 control and 5 applications. The control group was germinated with tap water, and the application groups were germinated with paraquat and two different doses of GTLE. Germination was carried out at room temperature for 72 h. At the end of the period, A. cepa bulbs were prepared for physiological, cytogenetic, biochemical, and anatomical analyzes using routine preparation techniques. As a result, paraquat application caused a decrease in physiological parameters and an increase in cytogenetic (except MI) and biochemical parameters. Compared to the control (group I), the germination percentage decreased by 38%, root length 12.5 times, and weight gain 5 times decreased in group IV treated with paraquat. MDA level increased 2.58 times, SOD activity 2.48 times, and CAT activity 4.51 times increased. Paraquat application caused a decrease in the percentage of MI and an increase in the number of MN and CAs. Paraquat application caused CAs in the form of fragment, sticky chromosome, unequal distribution of chromatin, bridge, nucleus with vacuoles, nucleus bud, and reverse polarization. In the meristematic cells of the root tips applied paraquat, unclearly vascular tissue, flattened cell nucleus, epidermis, and cortex cell deformation were observed. The application of GTLE together with paraquat caused an increase in the physiological parameter values and a decrease in the cytogenetic (except MI) and biochemical parameter values. An improvement in the severity of damages induced by paraquat was also observed in root tip meristematic cells. It was determined that the improvements observed in all these parameters were related to the dose of GTLE applied. The 380 mg/L dose of GTLE provided more protection than the 190 mg/L dose. Compared to group IV in which paraquat was applied, the germination percentage increased by 21%, root length 5.83 times, and weight gain 2.92 times increased in group VI administered 380 mg/L dose of GTLE. In addition, MDA level decreased 1.78 times, SOD activity 1.59 times and CAT activity 1.65 times. In conclusion, paraquat administration at a dose of 100 mg/L caused physiological, cytogenetic, biochemical, and anatomical toxicity in A. cepa bulbs. GTLE application, on the other hand, resulted in improvements in the severity of this toxicity induced by paraquat, depending on the dose. Therefore, GTLE can be used as an effective nutritional supplement to reduce or prevent the toxicity caused by environmental agents such as pesticides.
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