Towards water-free biobanks: long-term dry-preservation at room temperature of desiccation-sensitive enzyme luciferase in air-dried insect cells

Kikuta, Shingo; Watanabe, Shinya; Satō, Ryōichi; Gusev, Oleg; Несмелов, А. А.; Sogame, Yoichiro; Cornette, Richard; Kikawada, Takahiro

doi:10.1101/120063

Cited by 2 publications

(2 citation statements)

References 34 publications

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“…Na medicina, elementos-chave da anidrobiose poderiam ser aplicadas no armazenamento (i.e., "biobancos"), manutenção e transporte de estruturas biológicas, como células, sêmen (POLGE et al, 1949), sangue e até órgãos inteiros a seco (SARAGUSTY e LOI, 2019). Recentemente, um estudo apresentou a capacidade de preservação da enzima luciferase (Luc) no estado seco em até 372 dias, em temperatura ambiente, no interior de células Pv11 tolerantes à dessecação (KIKUTA et al, 2017).…”

Section: Agradeço Ao Departamento De Genética Da Faculdade De Medicinunclassified

Análises in silico de metalotioneínas (MTs) e a tolerância de Panagrolaimus superbus à hipóxia por imersão em matriz metálica de Gálio

Contiliani¹

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In silico analysis of metallothioneins (MTs) and the tolerance of Panagrolaimus superbus to hypoxia by immersion in a Gallium matrix. The free-living nematode Panagrolaimus superbus is able to enter into a state of suspended animation when exposed to extreme desiccation-phenomenon known as anhydrobiosis (life without water). During the dry state, anhydrobiotic organisms tolerate several physical stresses, such as extremes of temperature, vacuum and hyperacceleration. Metallothioneins (MTs) are ubiquitous proteins with short chains, capable of acting on detoxification and homeostasis of heavy metals due to the high content of cysteines in their chains. Considering that desiccated anhydrobiotic organisms remain viable for long periods in dry soils, in which high concentrations of metal ions are exhibited, we hypothesized that these organisms show tolerance to heavy metals due to structural differences in their MTs. Therefore, the objectives of this work were to investigate the tolerance of P. superbus to complete immersion in Gallium (Ga), by analyzing its viability and population growth, and to identify structural differences in MTs between anhydrobiotic and non-anhydrobiotic organisms by performing in silico analyses (cysteine composition and conservation, structural modeling, primary and tertiary structure analyses and alignment, and ligand affinity). The results demonstrate the high tolerance of P. superbus to hypoxic confinement in Ga for 7 days, with no negative effects on its viability and population growth. Additionally, the results of in silico analyses show higher values of cysteine composition and conservation in nonanhydrobiotic MTs, but no differences in tertiary structures and ligand interactions were shown. Thus, we report for the first time the preservation of an anhydrobiotic organism inside a heavy metal matrix. However, we suggest that this anhydrobiosis-mediated tolerance is not conferred by structural differences in MTs, but by their differential gene expression and/or involve a set of elements of the anhydrobiosis repair machinery.

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Section: Agradeço Ao Departamento De Genética Da Faculdade De Medicinunclassified

Análises in silico de metalotioneínas (MTs) e a tolerância de Panagrolaimus superbus à hipóxia por imersão em matriz metálica de Gálio

Contiliani¹

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“…Estudos recentes demonstraram a capacidade de preservação enzimática no estado seco por mais de 370 dias, em temperatura ambiente, no interior de células tolerantes a dessecação (Kikuta et al, 2017). Similarmente, o desenvolvimento de vacinas a seco capazes de serem armazenadas em temperatura ambiente também foi demonstrado (Alcock et al, 2010).…”

Section: Engenharia Anidrobióticaunclassified

Análises in silico de DAF-2 e exposição de Panagrolaimus superbus em câmara anóxica de nitrogênio (N2)

Ribeiro¹

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In silico analysis of DAF-2 and exposure of Panagrolaimus superbus in anoxic Nitrogen (N 2 ) chamber Anhydrobiosis, a term derived from the Greek "life without water", is a curious state of suspended animation into which certain organisms enter in response to extreme desiccation. Currently, organisms representing all kingdoms have already been identified with this ability, including the nematode used in this study, Panagrolaimus superbus. The anhydrobiotic state allows tolerance to a wide variety of physical stresses such as temperature extremes, hyperacceleration, and immersion in gallium. Due to the interesting properties that anhydrobiosis gives to organisms, a better understanding of this phenomenon has aroused the interest of the scientific community. In this sense, a central issue is the characterization of the ametabolic state, still little accepted by the scientific community. In parallel, the daf-2 gene encodes a protein that directs the development of the nematode Caenorhabditis elegans, its role being essential for anhydrobiosis.Considering the context, this study aims to contribute to a better definition of the metabolism of anhydrobiotic organisms and to investigate the protein encoded by daf-2 in search of possible anhydrobiosis-related molecular signatures. Thus, the anhydrobiont organism P. superbus was kept for one, three, and six months in an anoxic chamber, and we in silico analyzed (e.g., composition, structural modeling, analysis, and alignment of primary and tertiary structures) the DAF-2 protein in anhydrobiont organisms and nonanhydrobionts. The results found suggest that the protein of anhydrobiont organisms is significantly more basic, although it does not present divergences in its tertiary structures.We also demonstrated that the exposure of P. superbus desiccated in an anoxic N2 chamber for one, three, and six months, does not negatively impact its viability, when compared to their respective control group. Therefore, we add new evidence to the hypothesis of ametabolism during anhydrobiosis and we point out the basicity characteristic as something of importance for this interesting state.

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Towards water-free biobanks: long-term dry-preservation at room temperature of desiccation-sensitive enzyme luciferase in air-dried insect cells

Cited by 2 publications

References 34 publications

Análises in silico de metalotioneínas (MTs) e a tolerância de Panagrolaimus superbus à hipóxia por imersão em matriz metálica de Gálio

Análises in silico de metalotioneínas (MTs) e a tolerância de Panagrolaimus superbus à hipóxia por imersão em matriz metálica de Gálio

Análises in silico de DAF-2 e exposição de Panagrolaimus superbus em câmara anóxica de nitrogênio (N2)

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