Cytotoxic effect of aluminium ions on unicellular eukaryotic organism

Article Details: Received: 2019-10-08 | Accepted: 2019-11-26 | Available online: 2019-12-31https://doi.org/10.15414/afz.2019.22.04.130-137Aluminium is abundant in nature, food, or water and thus its exposition is part of everyday life. However, overexposure can result in cellular malfunctions. Therefore, the aim of this study was to investigate the effects of aluminium on eukaryotes, with the use of Schizosaccharomyces pombe as model organism. Spectrophotometry at OD600, inductively-coupled plasma optical emission spectroscopy (ICP-OES) and microscopy techniques were used to analyse aluminium responses on the living system. Our results revealed that exposition of increasing aluminium concentrations lead to cell growth inhibition in a concentration dependent manner. Furthermore, aluminium incorporation by the cell from media markedly increased with rising Al concentration. Our results indicate that the yeast self-protection system in the presence of lower Al(OH)3 concentration in the environment avoids to large extent dramatic uptake of aluminium by the cell while cells surrounded by higher aluminium concentrations lose this ability. Supplementation of the growth media with 100 μM Al(OH)3 doubled the amount of Al in the cell compared to untreated control (232 mg/kg vs. 459 mg/kg), whereas addition of 1 mM Al(OH)3 caused more than hundred fold increase of intracellular Al content (27,781 mg/kg). Here we also show that high concentrations of aluminium have an impact on cell morphology leading to cell integrity disruption. Findings presented in this study have the ambition to bring more light in an issue of how aluminium mediates impairments of the living organism.ReferencesANOOP, V.M. et al. (2003) Modulation of citrate metabolism alters aluminum tolerance in yeast and transgenic canola overexpressing a mitochondrial citrate synthase. In Plant Physiology, vol. 132, no. 4, pp. 2205–2217. DOI: https://doi.org/10.1104/pp.103.023903BRUNNER, I. and SPERISEN, C. (2013) Aluminum exclusion and aluminum tolerance in woody plants. In Frontiers in Plant  Science, vol. 4, 172. DOI: https://doi.org/10.3389/fpls.2013.00172CHEN, Y. et al. (2018) Advances in dialysis encephalopathy research: a review. In Neurological Sciences, vol. 39, no. 7, pp.  1151–1159. DOI: https://doi.org/10.1007/s10072-018-3426-yCLEMENS, S. and SIMM, C. (2003) Schizosaccharomyces pombe  as a model for metal homeostasis in plant cells: the phytochelatin‐dependent pathway is the main cadmium detoxification mechanism. In New Phytologist, vol. 159, no. 2, pp. 323–330. DOI: https://doi.org/10.1046/j.1469-8137.2003.00811.xCOLOMINA M.T. and PERIS-SAMPEDRO F. (2017) Aluminum and Alzheimer’s Disease. In Aschner M., Costa L. (eds.) Neurotoxicity of Metals. Advances in Neurobiology, vol.  18 Springer, Cham, pp. 183–197. DOI: https://doi. org/10.1007/978-3-319-60189-2_9FRANTZIOS, G. et al. (2000) Aluminium effects on microtubule organization in dividing root-tip cells of Triticum turgidum. I. Mitotic cells. In New Phytologist, vol. 145, no. 2, pp. 211–224. DOI:  https://doi.org/10.1046/j.1469-8137.2000.00580.xGUO, L. et al. (2016) Global fitness profiling identifies arsenic and cadmium tolerance mechanisms in fission yeast. In G3: Genes, Genomes, Genetics, vol. 6, no. 10, pp.3317–3333. DOI: https://doi.org./10.1534/g3.116.033829HEIMLICHER, M.B. et al. (2019) Reversible solidification of fission yeast cytoplasm after prolonged nutrient starvation. In Journal of Cell Science, vol.132, no. 21, pp. 1–17. DOI: https://doi.org/10.1242/jcs.231688HOFFMAN, C.S. et al. (2015) An ancient yeast for young geneticists: a primer on the Schizosaccharomyces pombe model system. In Genetics, vol. 201, no. 2, pp. 403–423. DOI: https://doi.org/10.1534/genetics.115.181503HUANG, W.-J. et al. (2014) Aluminum induces rapidly mitochondria-dependent programmed cell death in Alsensitive peanut root tips. In Botanical Studies, vol. 55, 67. DOI: https://doi.org/10.1186/s40529-014-0067-1JASKOWIAK, J. et al. (2018) Analysis of aluminum toxicity in Hordeum vulgare roots with an emphasis on DNA integrity and cell cycle. In PLoS ONE, vol. 13, 2: e0193156. DOI: https://doi.org/10.1371/journal.pone.0193156JONES, D. and RYAN, P. (2003) Aluminum toxicity. In: Thomas, B. et al. (eds.) Encyclopedia of Applied Plant Science. London: Elsevier Academic Press, pp. 656–664.KAIZER, R.R. et al. (2005) Acetylcholinesterase activation and enhanced lipid peroxidation after long-term exposure to low levels of aluminum on different mouse brain regions. In Journal of Inorganic Biochemistry, vol. 99, no. 9, pp. 1865–1870. DOI: https://doi.org/10.1016/j.jinorgbio.2005.06.015KAKIMOTO, M. et al. (2005) Genome-wide screening of aluminum tolerance in Saccharomyces cerevisiae. In Biometals: An International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine, vol. 18, no. 5, pp. 467–474. DOI: https://doi.org/10.1007/s10534-005-4663-0KOVACIK, A. et al. (2019) Trace metals in the freshwater fish  Cyprinus carpio: Effect to Serum Biochemistry and Oxidative  Status Markers. In Biological Trace Element Research, vol. 188, no. 2, pp. 494–507. DOI: https://doi.org/10.1007/ s12011-018-1415-xKUMAR, V. et al. (2009) Aluminium-induced oxidative DNA damage recognition and cellcycle disruption in different regions of rat brain. In Toxicology, vol. 264, no. 3, pp. 137–144. DOI: https://doi.org/10.1016/j.tox.2009.05.011LI, X. et al. (2011) Regulating cytoplasmic calcium homeostasis can reduce aluminum toxicity in yeast. In PLoS ONE, vol. 6, 6: e21148. DOI: https://doi.org/10.1371/journal.pone.0021148LI, X. et al. (2012) Aluminum induces osteoblast apoptosis through the oxidative stress-mediated JNK signaling pathway. In Biological Trace Element Research, vol. 150, no. 1, pp. 502–508. DOI: https://doi.org/10.1007/s12011-012-9523-5MACDIARMID, C.W. and Gardner, R.C. (1996) Al toxicity in yeast (A role for Mg?). In Plant Physiology, vol. 112, no. 3, pp. 1101–1109. DOI:  https://doi.org/10.1104/pp.112.3.1101MAYA, S. et al. (2016) Multifaceted effects of aluminium in neurodegenerative diseases: a  review. In Biomedicine & Pharmacotherapy, vol. 83, pp. 746–754. DOI: https://doi. org/10.1016/j.biopha.2016.07.035NEHRU, B. and ANAND, P. (2005) Oxidative damage following  chronic aluminium exposure in adult and pup rat brains. In Journal of Trace Elements in Medicine and Biology, vol. 19, no. 2, pp. 203–208. DOI: https://doi.org/10.1016/j.jtemb.2005.09.004NIAN, H. et al. (2012) Physiological and transcriptional analysis of the effects of aluminum stress on Cryptococcus humicola. In World Journal of Microbiology and Biotechnology, vol. 28, no. 6, pp. 2319–2329. DOI: https://doi.org/10.1007/ s11274-012-1039-9POZGAJOVA, M. et al. (2013) Prp4 kinase is required for proper segregation of chromosomes during meiosis in Schizosaccharomyces pombe. In Acta Biochimica Polonica, vol. 60, no. 4, pp. 871–873. DOI: https://doi.org/10.18388/ abp.2013_2075POZGAJOVA, M. et al.  (2019)  Impact of cadmium and nickel on ion homeostasis in the yeast  Schizosaccharomyces pombe.  In  Journal of Environmental Science and Health, Part  B,  pp. 1–8. DOI: https://doi.org/10.1080/03601234.2019.1673613PŘIBYL, P. et al. (2008) Cytoskeletal alterations in interphase  cells of the green alga Spirogyra decimina in response to heavy metals exposure: II. The effect of aluminium, nickel and copper. In Toxicology in Vitro, vol. 22, no. 5, pp. 1160– 1168. DOI: https://doi.org/10.1016/j.tiv.2008.03.005QIN, R. et al. (2010) Effects of aluminum on nucleoli in root tip cells and selected physiological and biochemical characters in Allium cepa var. agrogarum L. In BMC Plant Biology, vol. 10, p. 225. DOI: https://doi.org/10.1186/1471-2229-10-225SABATINOS, S.A. and FORSBURG, S.L. (2010) Molecular genetics of Schizosaccharomyces pombe. In Methods in Enzymology, vol. 470, pp. 759–795. DOI: https://doi.org/10.1016/S0076-6879(10)70032-XSALGADO, A. et al. (2012) Response to arsenate treatment in Schizosaccharomyces pombe and the role of its arsenate reductase activity. In PloS one, vol. 7, no. 8, pp.SHORT, A.I. et al. (1980) Reversible microcytic hypochromic anaemia in dialysis patients due to aluminium intoxication. In  Proceedings of the European Dialysis and Transplant Association. In European Dialysis and Transplant Association, vol. 17, pp. 226–233.TUN, N.M. et al. (2013) Disulfide stress-induced aluminium toxicity: molecular insights through genome-wide screening of  Saccharomyces cerevisiae. In Metallomics, vol. 5 no. 8, pp. 1068–1075. DOI: https://doi.org/10.1039/C3MT00083DVARDAR, F. et al. (2016) Determination of aluminum induced programmed cell death characterized by DNA fragmentation in Gramineae species. In Caryologia, vol. 69, no. 2, pp. 111–115. DOI: https://doi.org/10.1080/00087114.2015.1109954WANG, C. et al. (2013) Proteomic analysis of a high aluminum tolerant yeast Rhodotorula taiwanensis RS1 in response to aluminum stress. In Biochimica et Biophysica Acta (BBA) – Proteins and Proteomics, vol. 1834, no. 10, pp. 1969–1975. DOI: https://doi.org/10.1016/j.bbapap.2013.06.014WOOD, V. et al. (2002) The genome sequence of Schizosaccharomyces pombe. In Nature, vol. 415, no. 6874, pp. 871–880. DOI: https://doi.org/10.1038/nature724WU, M.J. et al. (2012) Delineation of the molecular mechanism for disulfide stress-induced aluminium toxicity. In Biometals: An International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine, vol. 25, no. 3, pp. 553–561. DOI: https://doi.org/10.1007/s10534-012-9534-xWYSOCKI, R., and TAMÁS, M. J. (2010) How Saccharomyces cerevisiae copes with toxic metals and metalloids. In FEMS microbiology reviews, vol. 34, no. 6, pp. 925–951. DOI: https://doi.org/10.1111/j.1574-6976.2010.00217.xZHANG, H. et al. (2014) Accumulation and cellular toxicity of aluminum in seedling of Pinus massoniana. In BMC Plant Biology,  vol. 14, 264. DOI: https://doi.org/10.1186/s12870-014-0264-9ZHENG, K. et al. (2007) Programmed cell death-involved aluminum toxicity in yeast alleviated by antiapoptotic members  with decreased calcium signals. In Plant Physiology, vol. 143, no. 1, pp. 38–49. DOI: https://doi.org/10.1104/ pp.106.08249

Aspirin Induces Platelet Receptor Shedding via ADAM17 (TACE)

Aspirin is effective in the therapy of cardiovascular diseases, because it causes acetylation of cyclooxygenase 1 (COX-1) leading to irreversible inhibition of platelets. Additional mechanisms can be suspected, because patients treated with other platelet COX inhibitors such as indomethacin do not display an increased bleeding tendency as observed for aspirin-treated patients. Recently, aspirin and other anti-inflammatory drugs were shown to induce shedding of L-selectin in neutrophils in a metalloproteinase-dependent manner. Therefore, we investigated the effects of aspirin on the von Willebrand Factor receptor complex glycoprotein (GP) Ib-V-IX, whose lack or dysfunction causes bleeding in patients. As quantified by fluorescence-activated cell sorting analysis in whole blood, aspirin, but not its metabolite salicylic acid, induced dose-dependent shedding of human and murine GPIbalpha and GPV from the platelet surface, whereas other glycoproteins remained unaffected by this treatment. Biotinylated fragments of GPV were detected by immunoprecipitation in the supernatant of washed mouse platelets, and the expression level of GPIbalpha was decreased in these platelets as measured by Western blot analysis. Although shedding occurred normally in COX-1-deficient murine platelets, shedding was completely blocked by a broad-range metalloproteinase inhibitor and, more importantly, in mouse platelets expressing an inactive form of ADAM17. Shed fragments of GPIbalpha and GPV were elevated in the plasma of aspirin-injected mice compared with animals injected with control buffer. These data demonstrate that aspirin at high concentrations induces shedding of GPIbalpha and GPV by an ADAM17-dependent mechanism and that this process can occur in vivo

Studien von formation und stabilizierung den pathologischen Thrombus in vivo

Platelet activation and adhesion resulting in thrombus growth is essential for normal hemostasis, but can lead to irreversible, life-threatening vessel occlusion. In the current study, the contribution of platelet integrins, activation receptors and the contact system of blood coagulation in such pathological conditions was investigated in mice.Plättchenaktivierung, -adhäsion und nachfolgende Thrombusbildung ist ein für die Hämostase essentieller Prozess, der jedoch zu irreversiblem lebensbedrohlichen Gefäßverschluss führen kann. In der vorliegenden Arbeit wurde die Rolle von Thrombozyten-Integrinen, aktivierenden Rezeptoren, sowie dem Kontaktsystem der Koagulation unter pathologischen Bedingungen im Maussystem untersucht

Prp4 kinase is required for proper segregation of chromosomes during meiosis in Schizosaccharomyces pombe

Chromosome segregation during meiosis is a complex process, which leads to production of four haploid gametes from two precursor cells. Reversible phosphorylation of proteins plays a crucial role in this process. The Schizosaccharomyces pombe Prp4 is an essential serine/threonine protein kinase, which belongs to the Clk/Sty family. To study the role of Prp4 in meiosis, we analysed chromosome segregation in a strain carrying conditional analog-sensitive allele of Prp4 protein kinase (prp4-as2). Our data show, that Prp4 protein kinase plays important role in chromosome segregation during meiosis, as revealed by enhanced missegregation of chromosomes in prp4-as2 mutant cells

Diminished thrombus formation and alleviation of myocardial infarction and reperfusion injury through antibody- or small-molecule-mediated inhibition of selectin-dependent platelet functions

BACKGROUND AND OBJECTIVES: P-selectin ctin has been implicated in important platelet functions. However, neither its role in thrombus formation and cardiovascular disorders nor its suitability as a therapeutic target structure is entirely clear. DESIGN AND METHODS: Platelet aggregation was assessed in complementary in vitro settings by measurements of static aggregation, standardized aggregometry and dynamic flow chamber assays. Degradation of aggregates was also analyzed under flow conditions using video microscopy. In vivo, platelet rolling in cutaneous venules was assessed by intravital microscopy in wild-type mice treated with selectin-blocking compounds as well as in P-selectin-deficient mice. FeCl3-induced arterial thrombosis was studied by intravital microscopy in untreated mice or mice treated with an inhibitor of selectin functions. Finally, inhibition of selectin functions was studied in an ischemia/reperfusion injury model in rats. RESULTS: Antibody- or small-molecule-mediated inhibition of P-selectin functions significantly diminished platelet aggregation (p<0.03) and platelet-neutrophil adhesion in vitro (p<0.01) as well as platelet aggregate sizes under flow (p<0.03). Established aggregates were degraded, either via detachment of single platelets following addition of efomycine M, or via detachment of multicellular clumps when P-selectin-directed Fab-fragments were used. In vivo, selectin inhibition resulted in a greater than 50% reduction of platelet rolling in cutaneous venules (p<0.01), producing rolling fractions similar to those observed in P-selectin-deficient mice (p<0.05). Moreover, inhibition of selectin functions significantly decreased the thrombus size in FeCl3-induced arterial thrombosis in mice (p<0.05). In an ischemia/reperfusion injury model in rats, small-molecule-mediated selectin inhibition significantly reduced myocardial infarct size from 18.9% to 9.42% (p<0.001) and reperfusion injury (p<0.001). INTERPRETATION AND CONCLUSIONS: Inhibition of P-selectin functions reduces platelet aggregation and can alleviate platelet-related disorders in disease-relevant preclinical settings