Role of Phytochelatins in Redox Caused Stress in Plants and Animals

Varied environmental compartments (such as soil and water) potentially contaminated with different metals/metalloids can impact the health of both plants and animals/humans. Trace amounts of Cu, Mn, Mo, Ni and Zn are beneficial for higher plants, whereas, Cr, Cu, Co, Mn, Mo, Se, V and Zn are known as the micronutrient metal/metalloids for animals/humans. However, elevated levels of the metals/metalloids can cause severe toxic consequences in both plants and animals/humans. Common in plants and animals/humans, phytochelatins (PCs), the principal non-protein, S-rich, thiolate peptides, protect (through different mechanisms) cellular functions and metal/metalloid homeostasis by performing their chelation and/or detoxification. With the major aim of broadening the current knowledge on the subject, this chapter (a) overviews PCs’ role and modulation separately in metal/metalloid-exposed plants and animals/humans; (b) discusses major methods for determination of PCs and bioassays for enzymes involved in PC synthesis; (c) evaluates the connection of PCs with bionanoparticles; and finally (d) highlights so far unexplored aspects in the present context

Microarray analysis of metallothioneins in human diseases: a review

K

MO38-2 Metallothionein-3: Potential therapeutic target for sorafenib resistance in hepatocellular carcinoma

1 p. JSMO2021 Virtual Congress. 2021 the Japanese Society of Medical Oncology Annual Meeting. February 18 - 21, 2021Background: Metallothionein-3 (M-3) has poorly characterized functions in hepatocellular carcinoma (HCC). HCC is a significant health problem. Globally is the second most common cause of cancer-associated death. Sorafenib was originally identified as an inhibitor of multiple oncogenic kinases and remains the only approved systemic therapy for advanced HCC. However, acquired resistance to sorafenib has been found in HCC patients, which results in poor prognosis. Overexpression of MT-3 decreased the sensitivity of HCC cells to sorefenib. Here, we investigated the impact of MT-3 up-regulation in HCC cells and the mechanisms underlying the sorafenib-resistance.Methods: To increase the expression of MT-3 HCC cells were transiently transfected with a plasmid containing MT-3 gene or with empty vector. The cDNA microarrays were accomplished using the ElectraSenseTM Reader. MS analysis was performed using a Q-Exactive MS. We used chick chorioallantoic membrane assay as in vivo model.Results: A cDNA profiling revealed that sorafenib resistance has a specific transcriptomic signature involving genes responsible for ion transport, trafficking and DNA repair. Also, The MS analysis data strongly suggest that resistance HCC cells acquired a complex regulatory network that significantly affects the ability of HCC cells to remove the ROS and activation of glycolysis. We provide the first evidence that up-regulation of MT3 resulted in increased dissociation, invasion, and intravasation from the primary tumours to the veins. In addition, MT3 profoundly impacted blood migration of Nbl cells and their extravasation to chicken organs.Conclusion: From a perspective of future utilization of our data, we anticipate that several identified genes and proteins could serve as prognostic biomarkers of outcome of sorafenib therapy. The increased expression of MT-3 within tumour mass should inform about worse prognosis and also decreased efficiency of sorafenib-based chemotherapy in HCC.Peer reviewe

GintGRX1, the first characterized glomeromycotan glutaredoxin, is a multifunctional enzyme that responds to oxidative stress

Glutaredoxins (GRXs) are small proteins with glutathione-dependent disulfide oxidoreductase activity involved in cellular defense against oxidative stress. This work reports the identification and characterization of the first glomeromycotan dithiol glutaredoxin gene from the fungus Glomus intraradices. The corresponding gene, named GintGRX1, shares high sequence similarity with previously described fungal GRXs. GintGRX1 contains the characteristic dithiol active site CPYC. By using a yeast expression system, we found that GintGRX1 encodes a multifunctional protein with oxidoreductase, peroxidase and glutathione S-transferase activity. GintGRX1 partially reverted sensitivity to superoxide radicals of the Δgrx1Δgrx2 Saccharomyces cerevisiae strain. GintGRX1 was transcriptionally regulated by paraquat but not by hydrogen peroxide. Copper induced an accumulation of reactive oxygen species in the extraradical mycelium of G. intraradices and up-regulation of GintGRX1 transcript levels. These data suggest a role for GintGRX1 in protecting the fungus against the oxidative damage induced directly by the superoxide anion or indirectly by copper. © 2008 Elsevier Inc. All rights reserved.This work was founded by a grant from the Consejería de Innovación, Ciencia y Empresa of the Junta de Andalucía, Spain (P06-CVI-02263). Karim Benabdellah was supported by an I3P contract from the Spanish Council for Scientific Research (CSIC). We would like to thank Drs. José Miguel Barea and Francisco Martín for helpful discussions and Ascención Valderas for excellent technical assistance.Peer Reviewe

The arbuscular mycorrhizal fungus Rhizophagus irregularis differentially regulates the copper response of two maize cultivars differing in copper tolerance

Arbuscular mycorrhiza can increase plant tolerance to heavy metals. The effects of arbuscular mycorrhiza on plant metal tolerance vary depending on the fungal and plant species involved. Here, we report the effect of the arbuscular mycorrhizal fungus Rhizophagus irregularis on the physiological and biochemical responses to Cu of two maize genotypes differing in Cu tolerance, the Cu-sensitive cv. Orense and the Cu-tolerant cv. Oropesa. Development of the symbiosis confers an increased Cu tolerance to cv. Orense. Root and shoot Cu concentrations were lower in mycorrhizal than in non-mycorrhizal plants of both cultivars. Shoot lipid peroxidation increased with soil Cu content only in non-mycorrhizal plants of the Cu-sensitive cultivar. Root lipid peroxidation increased with soil Cu content, except in mycorrhizal plants grown at 250 mg Cu kgsoil. In shoots of mycorrhizal plants of both cultivars, superoxide dismutase, ascorbate peroxidase, catalase and glutathione reductase activities were not affected by soil Cu content. In Cu-supplemented soils, total phytochelatin content increased in shoots of mycorrhizal cv. Orense but decreased in cv. Oropesa. Overall, these data suggest that the increased Cu tolerance of mycorrhizal plants of cv. Orense could be due to an increased induction of shoot phytochelatin biosynthesis by the symbiosis in this cultivar.Peer Reviewe

Proteomic analysis of UKF-NB-4 cells reveals a stimulatory activity of MT-3 on cellular senescence and apoptosis

1 p.Background: Metallothioneins (MTs) family is a intracellular and cysteine-rich proteins with a high affinity to metals. MT-3 could play important neuromodulatory and neuroprotective roles. MT-3 has been also found up-regulated in a number of cancers. Neuroblastoma (Nbl) is a cancer is the most common extra-cranial solid tumour of childhood. The main aim was to provide novel insights into the molecular mechanisms of hMT-3 up-regulation and to elucidate the effects beneath the MT-3 up-regulation in Nbl cells.Methods: To increase the expression of MT-3 Nbl (UKF-NB-4) cells were transiently transfected with a plasmid containing MT-3 gene (pcDNA3.1-GFP-hMT-3-TOPO). Separations of tryptic digests were carried out on an Easy-nLC 1000 nano system. MS analysis was performed using a Q-Exactive MS. The mass spectrum *.raw file was searched against the human SwissProt 57.15 database using MASCOT search engine (version 2.3, Matrix Science).Results: The efficiency of transfection analysed through a fluorescence of GFP tag expressed at the C-terminus of MT-3 showed more 70% transfection efficiency for both constructed plasmids (mock and MT-3). From the total of common proteins in dataset (hMT-3 vs. mock), 176, 20 and 1244 proteins were quantitatively identified with up-regulation, down-regulation, and no significant differences between hMT-3 and mock treatments. Noteworthy, the bioinformatical analyses revealed the exclusive expression (induced by MT-3) and up-regulation proteins of a number of proteins affecting biological pathways related to mitotic cell cycle, cellular responses to stress, positive regulation of proteolysis, negative regulation of cell cycle and programmed cell death.Conclusions: Our proteomic data shed some light on the proteins involved in inducing senescence and apoptosis in tested Nbl cells with up-regulated MT-3. Organisms with renewable tissues had to evolve mechanisms to prevent the development of cancer. Cellular senescence and apoptosis are among those mechanisms. Further experiments will be performed to functionally verify these data to provide novel insights into the Nbl biology. These might be useful to develop novel therapeutic protocols utilizing MT-3 as prognostic biomarker or therapeutic target.European Research Council (ERC) under the European Uniońs Horizon 2020 Research and Innovation Programme (grant agreement No. 759585).Peer reviewe

Vývoj magnetického elektrochemického barkodového pole pro analýzu bodových mutací v genu neuraminidazy H5N1

Since its first official detection in the Guangdong province of China in 1996, the highly pathogenic avian influenza virus of H5N1 subtype (HPAI H5N1) has reportedly been the cause of outbreaks in birds in more than 60 countries, 24 of which were European. The main issue is still to develop effective antiviral drugs. In this case, single point mutation in the neuraminidase gene, which causes resistance to antiviral drug and is, therefore, subjected to many studies including ours, was observed. In this study, we developed magnetic electrochemical bar code array for detection of single point mutations (mismatches in up to four nucleotides) in H5N1 neuraminidase gene. Paramagnetic particles Dynabeads with covalently bound oligo (dT)25 were used as a tool for isolation of complementary H5N1 chains (H5N1 Zhejin, China and Aichi). For detection of H5N1 chains, oligonucleotide chains of lengths of 12 (+5 adenine) or 28 (+5 adenine) bp labeled with quantum dots (CdS, ZnS and/or PbS) were used. Individual probes hybridized to target molecules specifically with efficiency higher than 60%. The obtained signals identified mutations present in the sequence. Suggested experimental procedure allows obtaining further information from the redox signals of nucleic acids. Moreover, the used biosensor exhibits sequence specificity and low limits of detection of subnanogram quantities of target nucleic acidsOd svého prvního oficiálního odhalení v provincii Guangdong v Číně v roce 1996, byl vysoce patogenní virus influenzy ptáků podtypu H5N1 (HPAI H5N1) údajně příčinou vyvolání choroby u ptáků ve více než 60 zemích světa, z nichž 24 bylo evropských. Hlavním problémem je i nadále rozvíjení efektivních antivirotik. V tomto případě byla pozorována jediná bodová mutace v genu neuraminidázy, která způsobuje odolnost vůči antivirovému léku, a je proto podroben mnoha studií, včetně té naší. V této studii jsme vyvinuli elektrochemické barkodové pole pro detekci jednotlivých bodových mutací (nesoulad až do čtyř nukleotidů) v genu neuraminidázy H5N1. Paramagnetické částice Dynabeads s kovalentně vázaným oligo dT (25) byly použity jako nástroje pro izolaci komplementárních řetězců H5N1 (H5N1 Zhejin, Čína a Aichi). Pro detekci řetězců H5N1 byly použity řetězce oligonukleotidů o délce 12 (5 adenin), nebo 28 (5 adenin) bp označené kvantovými tečkami (CdS, ZnS a / nebo PbS). Jednotlivé sondy se hybridizují k cílovým molekulám specificky s účinností vyšší než 60 %. Získané signály identifikovaly přítomnost mutací v sekvencích. Doporučený experimentální postup umožňuje získat další informace redoxních signálů nukleových kyselin. Použitý biosensor vykazuje sekvenční specificitu a nízké meze detekce v subnanogramových množství cílové nukleové kyseliny

Correction: The Zinc-Schiff Base-Novicidin Complex as a Potential Prostate Cancer Therapy.

[This corrects the article DOI: 10.1371/journal.pone.0163983.]