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1.
Appl Environ Microbiol ; 88(1): e0176021, 2022 01 11.
Artigo em Inglês | MEDLINE | ID: mdl-34669425

RESUMO

Frequently, laccases are triggered during fungal cocultivation for overexpression. The function of these activated laccases during coculture has not been clarified. Previously, we reported that Gongronella sp. w5 (w5) (Mucoromycota, Mucoromycetes) specifically triggered the laccase Lcc9 overexpression in Coprinopsis cinerea (Basidiomycota, Agaricomycetes). To systematically analyze the function of the overexpressed laccase during fungal interaction, C. cinerea mycelia before and after the initial Lcc9 overexpression were chosen for transcriptome analysis. Results showed that accompanied by specific utilization of fructose as carbohydrate substrate, oxidative stress derived from antagonistic compounds secreted by w5 appears to be a signal critical for laccase production in C. cinerea. A decrease in reactive oxygen species (ROS) in the C. cinerea wild-type strain followed the increase in laccase production, and then lcc9 transcription and laccase activity stopped. By comparison, increased H2O2 content and mycelial ROS levels were observed during the entire cocultivation in lcc9 silenced C. cinerea strains. Moreover, lcc9 silencing slowed down the C. cinerea mycelial growth, affected hyphal morphology, and decreased the asexual sporulation in coculture. Our results showed that intracellular ROS acted as signal molecules to stimulate defense responses by C. cinerea with the expression of oxidative stress response regulator Skn7 and various detoxification proteins. Lcc9 takes part in a defense strategy to eliminate oxidative stress during the interspecific interaction with w5. IMPORTANCE The overproduction of laccase during interspecific fungal interactions is well known. However, the exact role of the upregulated laccases remains underexplored. Based on comparative transcriptomic analysis of C. cinerea and gene silencing of laccase Lcc9, here we show that oxidative stress derived from antagonistic compounds secreted by Gongronella sp. w5 was a signal critical for laccase Lcc9 production in Coprinopsis cinerea. Intracellular ROS acted as signal molecules to stimulate defense responses by C. cinerea with the expression of oxidative stress response regulator Skn7 and various detoxification proteins. Ultimately, Lcc9 takes part in a defense strategy to eliminate oxidative stress and help cell growth and development during the interspecific interaction with Gongronella sp. w5. These findings deepened our understanding of fungal interactions in their natural population and communities.


Assuntos
Agaricales , Lacase , Agaricales/metabolismo , Proteínas Fúngicas/genética , Peróxido de Hidrogênio , Lacase/genética , Lacase/metabolismo , Estresse Oxidativo
2.
Ecotoxicol Environ Saf ; 208: 111603, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396123

RESUMO

Chlorothalonil (CLT) is a broad spectrum, and non-systemic fungicide applied in foliar structures to prevent and treat pathogens. This compound reaches to aquatic environments and affects the biota. In this context, the main goal of this study was to assess the effects of CLT at biochemical, tissular, and individual levels of biological organization using the invasive bivalve Corbicula largillierti as a bioindicator species. Clams were exposed to different sublethal concentrations (0, 10, 20 and 50 µg. L-1 CLT) for 96 h. At biochemical level, the enzymatic activity (Glutathione-s-Transferase, Catalase, Acetyl-, Butiryl- and Carboxyl-esterases) and lipid peroxidation were measured in gills and the visceral mass. Also, the digestive gland morphometry through quantitative histological indexes was registered at the tissular level. Finally, filtering activity and burial behavior at the individual level were measured. At the highest CLT concentration, the most significant changes were observed in enzymatic activity (except for butyrylcholinesterase), lipid peroxidation and in digestive gland morphometry. It was also registered increases of the filtering activity and the latency time to burial. Most of the biomarkers assessed showed significant responses under CLT exposure. Therefore, taking into account that C. largillierti was affected by CLT, it can be expected that other species could be in a potential risk if this fungicide is present in freshwater systems.


Assuntos
Corbicula/efeitos dos fármacos , Monitoramento Ambiental/métodos , Água Doce/química , Fungicidas Industriais/toxicidade , Nitrilas/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Biomarcadores/metabolismo , Catalase/metabolismo , Corbicula/enzimologia , Relação Dose-Resposta a Droga , Fungicidas Industriais/análise , Brânquias/efeitos dos fármacos , Brânquias/enzimologia , Glutationa Transferase/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Nitrilas/análise , Poluentes Químicos da Água/análise
3.
J Cell Biochem ; 120(2): 1362-1375, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30368880

RESUMO

The aim of this study was to investigate the role of NYD-SP15 in the growth and oxidative-stress responses of ARPE-19 cells. ARPE-19 cell lines overexpressing wild type or RNA interference against NYD-SP15 were established via lentivirus transfection. Cell growth and proliferation, migration, apoptosis, and cell cycle progression were monitored using the Cell Counting Kit-8 assay, the wound scratch assay, and flow cytometry, respectively. Caspase-3/8/9 activity was examined using the caspase-3/8/9 assay kit. An hydrogen peroxide (H 2 O 2 )-induced oxidative-stress damage model was used to study the effect of NYD-SP15 knockdown by examining the activity of reactive oxygen species (ROS). Expressions of Kelch-like ECH-associated protein 1 (Keap-1)/heme oxygenase-1 (HO-1)/nuclear factor erythroid 2-related factor 2 (Nrf2), mitogen-activated protein kinase (MAPK), and Akt were detected by Western blot analysis. The mRNA chip of NYD-SP15 overexpressed ARPE-19 cells as well as controls were performed by one array plus process. Overexpression (OE) of NYD-SP15 inhibited the proliferation and migration of ARPE-19 cells, and led to apoptosis and caspase-3/9 activation. OE of NYD-SP15 inhibited MAPKs and Akt signaling. Downregulation of NYD-SP15 had no effect on the growth of normally cultured ARP19 cells with 10% fetal bovine serum, but promoted the growth of ARP19 cells in the presence of starvation challenge. Gene chip showed that OE of NYD-SP15 led to downregulation of 254 genes and upregulation of 57 genes. Downregulation of NYD-SP15 also exerted a protective effect on H 2 O 2 -induced cell apoptosis and ROS. NYD-SP15 downregulation led to increments in the expression of Nrf2, Keap-1, and HO-1 in response to 200 µM H 2 O 2 . NYD-SP15 might inhibit the growth, proliferation, and migration and promote apoptosis of ARPE-19 cells via MAPK and Akt signaling. Downregulation of NYD-SP15 could protect ARPE-19 cells from H 2 O 2 -induced oxidative damage by active Keap-1/HO-1/Nrf2, Akt, and MAPK signaling.

4.
Proc Biol Sci ; 285(1883)2018 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-30051844

RESUMO

Natural and human-induced stressors elicit changes in energy metabolism and stress physiology in populations of a wide array of species. Cities are stressful environments that may lead to differential selection on stress-coping mechanisms. Given that city ponds are exposed to the urban heat island effect and receive polluted run-off, organisms inhabiting these ecosystems might show genetic differentiation for physiological traits enabling them to better cope with higher overall stress levels. A common garden study with 62 Daphnia magna genotypes from replicated urban and rural populations revealed that urban Daphnia have significantly higher concentrations of total body fat, proteins and sugars. Baseline activity levels of the antioxidant defence enzymes superoxide dismutase (SOD) and glutathione-S-transferase (GST) were higher in rural compared with city populations, yet urban animals were equally well protected against lipid peroxidation. Our results add to the recent evidence of urbanization-driven changes in stress physiology and energy metabolism in terrestrial organisms. Combining our results with data on urban life history evolution in Daphnia revealed that urban genotypes show a structured pace-of-life syndrome involving both life-history and physiological traits, whereas this is absent in rural populations.


Assuntos
Daphnia/fisiologia , Metabolismo Energético/genética , Variação Genética , Estresse Oxidativo/genética , Animais , Bélgica , Daphnia/genética , Características de História de Vida , Modelos Biológicos , Temperatura , Urbanização
5.
Appl Microbiol Biotechnol ; 101(18): 7027-7039, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28776099

RESUMO

On a daily basis, humans, and their colonizing microbiome, are exposed to both indoor and outdoor dust, containing both deleterious organic and inorganic contaminants, through dermal contact, inhalation, and ingestion. Recent studies evaluating the dust exposure responses of opportunistic pathogens, such as Escherichia coli and Pseudomonas aeruginosa, revealed significant increases in biofilm formation following dust exposure. In this study, the effects of dust exposure on mixed bacterial cultures as well as HT-29 co-cultures were evaluated. As it was observed in pure, single bacterial cultures earlier, neither indoor nor outdoor dust exposure (at concentrations of 100 µg/mL) influenced the growth of mixed bacterial liquid cultures. However, when in paired mixed cultures, dust exposure increased sensitivity to oxidative stress and significantly enhanced biofilm formation (outdoor dust). More specifically, mixed cultures (E. coli-Klebsiella pneumoniae, K. pneumoniae-P. aeruginosa, and E. coli-P. aeruginosa) exhibited increased sensitivity to 20 and 50 mM of H2O2 in comparison to their pure, single bacterial culture counterparts and significantly enhanced biofilm production for each mixed culture. Finally, bacterial proliferation during a eukaryotic gut cell (HT29) co-culture was significantly more robust for both K. pneumoniae and P. aeruginosa when exposed to both house and road dust; however, E. coli only experienced significantly enhanced proliferation, in HT29 co-culture, when exposed to road dust. Taken together, our findings demonstrate that bacteria respond to dust exposure differently when in the presence of multiple bacterial species or when in the presence of human gut epithelial cells, than when grown in isolation.


Assuntos
Biofilmes/crescimento & desenvolvimento , Poeira/análise , Escherichia coli/fisiologia , Klebsiella pneumoniae/fisiologia , Microbiota , Pseudomonas aeruginosa/fisiologia , Técnicas de Cocultura , Exposição Ambiental , Microbiologia Ambiental , Trato Gastrointestinal/microbiologia , Células HT29 , Humanos , Peróxido de Hidrogênio/farmacologia , Estresse Oxidativo
6.
Toxicol Mech Methods ; 26(4): 295-300, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27117342

RESUMO

Bisphenol A (BPA) is regularly detected in aquatic ecosystems due to increased use of products based on polycarbonate plastics and epoxy resins. It migrates from these products directly into rivers and marine waters or indirectly through effluents from wastewater treatment plants and landfilled sites. BPA can affect aquatic organisms both chronically and acutely at sensitive live stages. Despite reports indicating harmful effects of BPA, little is known about its role in oxidative stress responses in fish. In this study, we investigated the transcriptional effect of BPA (0, 1, 10, 100 µM) on an Atlantic salmon kidney (ASK) cell line for 6 h and 24 h by monitoring expression of 11 genes: elongation factor 1-alpha (ef1a), 18S ribosomal RNA (18s), gluthation (gsh), superoxide dismutase (sod), thioredoxin (txd), Salmo salar oxidative stress-responsive serine-rich 1 (oxr), glucose-regulated protein 78 (grp78), heat shock protein 70 (hsp70), sequestosome1 (p62), interleukin-1 beta (il-1beta) and toll-like receptor 8 (tlr8). In general, only the 100 µM concentration treatment altered the mRNA expression. BPA down-regulated the expression of gsh and sod genes for both exposure-times while txd gene was the only down-regulated after 6-h exposure. The up-regulation of genes in the ASK cell line exposed for 6 h was only observed in il-1beta, while the 24-h exposure resulted in the up-regulation of oxr, tlr8, hsp70, p62 and il-1beta genes. The last three genes increased several fold compared to the others. The results showed that BPA exposure at 100 µM imposed oxidative stress on the ASK cell line and longer exposure time involved transcriptional responses of immune-related genes. This may indicate the possible role of BPA-associated oxidative stress in induction of inflammatory response in this macrophage-like cell type.


Assuntos
Compostos Benzidrílicos/toxicidade , Rim/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fenóis/toxicidade , Salmo salar , Transcrição Gênica/genética , Poluentes Químicos da Água/toxicidade , Animais , Técnicas de Cultura de Células , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Rim/citologia , Rim/metabolismo , Estresse Oxidativo/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Tempo
7.
Antioxidants (Basel) ; 12(11)2023 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-38001754

RESUMO

Cyanobacterial blooms (CYBs) have become a global environmental issue, posing risks to edible bivalves. Toxic cyanobacteria and thermal stress represent the two key co-occurring stressors to bivalves experiencing CYBs. To investigate the combined effects of these stressors on the edible bivalve Corbicula fluminea, the responses to oxidative stress and the molecular mechanisms of physiological adaptations in C. fluminea were examined under co-exposure to toxic Microcystis aeruginosa and thermal stress. The activity of antioxidant enzymes, including GST, SOD, CAT, GPx and GR, was significantly influenced by the interaction between temperature and M. aeruginosa (p < 0.05). A positive correlation was observed between toxic M. aeruginosa exposure and elevated SOD and GPx activities at 30 °C, demonstrating that SOD and GPx may help C. fluminea defend effectively against MCs under thermal stress. Furthermore, significant interactive effects between toxic M. aeruginosa and temperature were also observed in ROS and MDA (p < 0.05). The results of the PCA and IBR index also evidenced the apparent influence of toxic M. aeruginosa and thermal stress on oxidative stress responses of C. fluminea. The eggNOG and GO annotations confirmed that a substantial portion of differentially expressed genes (DEGs) exhibited associations with responses to oxidative stress and transporter activity. Additionally, KEGG analysis revealed that abundant DEGs were involved in pathways related to inflammatory responses, immune functions and metabolic functions. These findings improve our understanding of the mechanism of the physiological adaptation in bivalves in response to cyanotoxins under thermal conditions, potentially enabling the evaluation of the viability of using bivalves as a bioremediation tool to manage CYBs in eutrophic waters.

8.
Environ Pollut ; 334: 122152, 2023 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-37414119

RESUMO

Ethiprole, a phenylpyrazole insecticide, has been increasingly used in the Neotropical region to control stink bug pests in soybean and maize fields. However, such abrupt increases in use may have unintended effects on non-target organisms, including those inhabiting freshwater ecosystems. Here, we evaluated the effects of acute (96 h) sublethal exposure to ethiprole (up to 180 µg/L, which is equivalent to 0.013% of the recommended field dose) on biomarkers of stress in the gills, liver, and muscle of the Neotropical fish Astyanax altiparanae. We further recorded potential ethiprole-induced effects on the structural histology of A. altiparanae gills and liver. Our results showed that ethiprole exposure increased glucose and cortisol levels in a concentration-dependent manner. Ethiprole-exposed fish also exhibited higher levels of malondialdehyde and greater activity of antioxidant enzymes, such as glutathione-S-transferase and catalase, in both gills and liver. Furthermore, ethiprole exposure led to increased catalase activity and carbonylated protein levels in muscle. Morphometric and pathological analyses of the gills revealed that increasing ethiprole concentration resulted in hyperemia and loss of integrity of the secondary lamellae. Similarly, histopathological analysis of the liver demonstrated higher prevalence of necrosis and inflammatory infiltrates with increasing ethiprole concentration. Altogether, our findings demonstrated that sublethal exposure to ethiprole can trigger a stress response in non-target fish species, which may lead to potential ecological and economic imbalances in Neotropical freshwater systems.


Assuntos
Characidae , Poluentes Químicos da Água , Animais , Catalase/metabolismo , Ecossistema , Estresse Oxidativo , Poluentes Químicos da Água/metabolismo , Antioxidantes/metabolismo , Glutationa Transferase/metabolismo , Fígado/metabolismo , Brânquias/metabolismo , Peroxidação de Lipídeos
9.
Front Physiol ; 13: 932537, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36117701

RESUMO

Freshwater pollution is a complex mixture of xenobiotics due to the wastewater and the various chemicals routinely applied to agricultural lands that are discharged into water bodies. Xenobiotics can exert damage to the aquatic biota threatening the biodiversity of aquatic ecosystems. The oxidative damage and antioxidant responses have been widely investigated in freshwater organisms, mainly in fish and some invertebrates but in freshwater snails are scarce. This study aimed to assess the oxidative stress exerted by potential toxicity of water from two rivers of the Mexican Atlantic Slope (Tecolutla and Tuxpan rivers) in a freshwater mollusk Physella acuta. Lipid peroxidation level and a battery of antioxidant enzymes (Superoxide dismutase, Catalase and Glutathione peroxidase) were measured in P. acuta. The results are contextualized from an ecological point of view, associating the bioassay results with water quality characteristics. Water samples were obtained from three study sites for each river (in two seasons: Northern wind and dry). Twelve water quality variables were analyzed, and an additional water sample was used to perform a static bioassay for 96 h with snails grown in laboratory. After the exposure, we assessed lipid peroxidation level and the antioxidant responses of P. acuta exposed to water of rivers, and the Integrated Biomarker Response was computed. The highest lipid peroxidation level occurred in organisms exposed to water during the Northern wind season in both rivers. During this season, in the Tecolutla river, the superoxide dismutase activity was able to counteract the lipid peroxidation process, representing an adaptive response. In contrast, in the Tuxpan river, the superoxide dismutase was unable to counteract that process, stimulating CAT and GPx activities. The Integrated Biomarker Response showed that the Tecolutla river had higher values in the upper reaches than the Tuxpan river, showing a decreasing downstream gradient in both seasons. In the Tuxpan river, during the Dry season, the IBR score showed an increasing downstream gradient. During the Northern wind season, the IBR was higher in the upper reaches of both rivers, possibly due to the increased materials transported by runoff from the catchment, which includes a complex mixture of xenobiotics that affects the health of the sentinel species and aquatic biota in general. Based on our results, Physella acuta is proposed as sentinel species.

10.
Am J Transl Res ; 13(5): 4796-4803, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34150060

RESUMO

OBJECTIVE: This paper aims to explore the effects of remifentanil combined with propofol on the stress responses, oxidative damage, and inflammatory responses in cardiac surgery patients. METHODS: One hundred and four patients who underwent cardiac surgery in our hospital from August 2017 to March 2019, were recruited as the study cohort and divided into control and observation groups. The 50 patients in the control group were anesthetized with fentanyl and propofol, and the 54 patients in the observation group were anesthetized with remifentanil and propofol. The general clinical data were observed and compared between the two groups. At different time points, changes in the oxidative stress response indicators (mean artery pressure (MAP) and heart rate (HR)) and in the cardiac function indexes (left ventricular ejection fraction (LVEF), stroke volume (SV), and cardiac output (CO)) were observed. The inflammatory cytokine levels (high-sensitivity C-reactive protein (hs-CRP), interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α)) were analyzed using enzyme-linked immunosorbent assays (ELISA). The patients' postoperative recovery (time to spontaneous respiration, time to opening eyes, extubation time) and their Visual Analogue Scale (VAS) scores were observed. Their pain at half an hour and at 24 hours after the operation were observed, as well as their postoperative adverse reactions. RESULTS: There were no differences in the general data between the two groups (P>0.05). Compared with the patients in the control group, the patients in the observation group had better oxidative stress levels and better cardiac function indexes (P<0.05), better postoperative inflammatory cytokine levels (P<0.05), better postoperative recovery (P<0.05), lower postoperative pain scores (P<0.05), and a lower total incidence of adverse reactions (P<0.05). CONCLUSION: Remifentanil combined with propofol can effectively reduce oxidative stress and inflammatory responses in cardiac surgery patients.

11.
Environ Int ; 157: 106867, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34537519

RESUMO

Passive equilibrium sampling of chemical mixtures from different human post-mortem tissues (liver, brain (cerebrum and cerebellum), adipose tissue) and blood was combined with instrumental analysis using direct sample introduction (DSI) GC-MS/MS and bioanalytical profiling using in vitro bioassays targeting the activation of the aryl hydrocarbon receptor (AhR-CALUX), the adaptive stress response (AREc32) and cytotoxicity. The tissues stemmed from pathology samples collected in two German cities and covered males and females aged 21 to 100 with a mean age of 67 years. Neutral organic chemicals were extracted using polydimethylsiloxane (PDMS) at mass ratios of tissue to PDMS of approximately 6 for blood, 3 for adipose tissue and 10 for liver and brain. Amounts of chemicals in PDMS were converted to lipid-associated concentrations using previously measured partition constants that were chemical-independent despite covering eight orders of magnitude in hydrophobicity. Up to 35 of 99 targeted chemicals were detected in 6 tissues of 16 individuals (88 samples in total), among them legacy persistent organic pollutants (POP) such as DDT and derivatives and polychlorinated biphenyls (PCB) but also modern pesticides and chemicals present in consumer products. POPs were highest in adipose tissue and lipid-associated concentrations increased with age, while concentrations of fragrance materials such as galaxolide were independent of age. In tissues from the same individual, chemical concentrations mostly increased from similar levels in brain and blood to higher levels in liver and highest in adipose tissue. However, easily degradable chemicals such as phenanthrene were mainly detected in blood and brain, and very hydrophilic chemicals were least abundant in adipose tissue. The passive sampling method allows a direct comparison of chemical burden between different tissues and may have forensic applications, for example to study internal distributions or to use one tissue type as a proxy for others. The sum of concentrations of the detected chemicals was positively correlated with the bioassay responses but mixture modeling showed that the detected chemicals explained less than 2% of the activation of the AhR and less than 0.5% of cytotoxicity. This means that more than 10,000 chemicals would need to be included in an analytical method to capture all the effects with many chemicals potentially being below detection limits but still contributing to mixture effects. Therefore, we propose a smart combination of chemical analysis and bioassays to quantify priority chemicals but use bioassay responses as effect-scaled concentrations to capture the entire exposome in future epidemiological studies.


Assuntos
Bifenilos Policlorados , Poluentes Químicos da Água , Idoso , Bioensaio , Feminino , Humanos , Masculino , Compostos Orgânicos , Silicones , Espectrometria de Massas em Tandem , Poluentes Químicos da Água/análise
12.
Aquat Toxicol ; 228: 105618, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32937231

RESUMO

Bifenthrin is a second generation synthetic pyrethroid insecticide that is widely used in Australia and worldwide. It is frequently found in urban freshwater sediments at concentrations likely to impact biota as it is highly toxic to fish and macroinvertebrates, such as chironomids. Our main goal was to evaluate if oxidative stress and hydrolase enzymes are useful biomarkers of effect of synthetic pyrethroids exposure under different scenarios. Chironomus tepperi larvae (5 days old) were exposed to sub-lethal sediment concentrations of bifenthrin for 5 days under controlled laboratory conditions. A field-based microcosm exposure with bifenthrin-spiked sediments (using the same concentrations as the laboratory exposure) was carried out at a clean field site for four weeks to allow for colonization and development of resident chironomid larvae. At the end of both experiments, Chironomus larvae (C. tepperi in the laboratory exposures and C. oppositus in the microcosm exposures) were collected and oxidative stress enzymes (Glutathione-s-Transferase, Glutathione Reductase and Glutathione Peroxidase) and hydrolase enzymes (Acetylcholinesterase and Carboxylesterase) were measured. Only the Glutathione Peroxidase activity was significantly impacted in larvae from the laboratory exposure. On the contrary, significant changes were observed in all the measured enzymes from the field-based microcosm exposure. This is likely because exposure was throughout the whole life cycle, from egg mass to fourth instar, showing a more realistic exposure scenario. Furthermore, this is the first time that changes in oxidative stress and hydrolase enzymes have been shown to occur in Australian non-biting midges exposed under field-based microcosm conditions. Thus, this study demonstrated the usefulness of these enzymes as biomarkers of effect following bifenthrin exposure in microcosms. It also highlights the importance of using a range of different biochemical endpoints to get a more holistic understanding of pesticide effects and the pathways involved.


Assuntos
Monitoramento Biológico/métodos , Chironomidae/efeitos dos fármacos , Larva/efeitos dos fármacos , Piretrinas/toxicidade , Poluentes Químicos da Água/toxicidade , Acetilcolinesterase/metabolismo , Animais , Austrália , Biomarcadores/metabolismo , Chironomidae/enzimologia , Sedimentos Geológicos/química , Glutationa Transferase/metabolismo , Larva/enzimologia , Estágios do Ciclo de Vida , Estresse Oxidativo/efeitos dos fármacos
13.
Front Microbiol ; 11: 590851, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33362738

RESUMO

NprR is a protein of Bacillus anthracis that exhibits moonlighting functions as either a phosphatase or a neutral protease regulator that belongs to the RNPP family. We previously observed that the extracellular protease activity of an nprR deletion mutant significantly decreased within in vitro cultures. To identify the genes within the regulatory network of nprR that contribute to its protease activity, integrated transcriptomic and proteomic analyses were conducted here by comparing the nprR deletion mutant and parent strains. A total of 366 differentially expressed genes (DEGs) between the strains were observed via RNA-seq analysis. In addition, label-free LC-MS/MS analysis revealed 503 differentially expressed proteins (DEPs) within the intracellular protein fraction and 213 extracellular DEPs with significant expressional differences between the strains. The majority of DEGs and DEPs were involved in environmental information processing and metabolism. Integrated transcriptomic and proteomic analyses indicated that oxidation-reduction-related GO terms for intracellular DEPs and endopeptidase-related GO terms for extracellular DEPs were significantly enriched in the mutant strain. Notably, many genes involved in protease activity were largely downregulated in the nprR deletion mutant cultures. Moreover, western blot analysis revealed that the major extracellular neutral protease Npr599 was barely expressed in the nprR deletion mutant strain. The mutant also exhibited impaired degradation of protective antigen, which is a major B. anthracis toxin component, thereby resulting in higher protein yields. Concomitantly, another global transcriptional regulator, SpxA1, was also dramatically downregulated in the nprR deletion mutant, resulting in higher sensitivity to oxidative and disulfide stress. These data consequently indicate that NprR is a transcriptional regulator that controls genes whose products function as extracellular proteases and also is involved in oxidative stress responses. This study thus contributes to a more comprehensive understanding of the biological function of NprR, and especially in the middle growth stages of B. anthracis.

14.
Aquat Toxicol ; 228: 105619, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32937230

RESUMO

Commonly affected by changes in climate and environmental conditions, coastal areas are very dynamic environments where shellfish play an important ecological role. In this study, the oxidative stress and genotoxic responses of mussels (Mytilus galloprovincialis) exposed to paralytic shellfish toxin (PST) - producing dinoflagellates Gymnodinium catenatum were evaluated under i) current conditions (CC: 19 °C; pH 8.0), ii) warming (W: 24 °C; pH 8.0), iii) acidification (A:19 °C; pH 7.6) and iv) combined effect of warming and acidification (WA: 24 °C; pH 7.6). Mussels were fed with G. catenatum for 5 days, and to a non-toxic diet during the following 10 days. A battery of oxidative stress biomarkers and comet assay was performed at the peak of toxin accumulation and at the end of the post-exposure phase. Under CC, gills and hepatopancreas displayed different responses/vulnerabilities and mechanisms to cope with PST. While gills presented a tendency for lipid peroxidation (LPO) and genetic damage (expressed by the Genetic Damage Indicator - GDI), hepatopancreas seems to better cope with the toxins, as no LPO was observed. However, the mechanisms involved in hepatopancreas protection were not enough to maintain DNA integrity. The absence of LPO, and the antioxidant system low responsiveness, suggests DNA damage was not oxidative. When exposed to toxic algae under W, toxin-modulated antioxidant responses were observed in both gills and hepatopancreas. Simultaneous exposure to the stressors highlighted gills susceptibility with a synergistic interaction increasing DNA damage. Exposure to toxic algae under A led to genotoxicity potentiation in both organs. The combined effect of WA did not cause relevant interactions in gills antioxidant responses, but stressors interactions impacted LPO and GDI. Antioxidant responses and LPO pointed out to be modulated by the environmental conditions in hepatopancreas, while GDI results support the dominance of toxin-triggered process. Overall, these results reveal that simultaneous exposure to warming, acidification and PSTs impairs mussel DNA integrity, compromising the genetic information due to the synergetic effects. Finally, this study highlights the increasing ecological risk of harmful algal blooms to Mytilus galloprovinciallis populations.


Assuntos
Dano ao DNA , Toxinas Marinhas/toxicidade , Mytilus/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Ensaio Cometa , Dinoflagellida/metabolismo , Brânquias/efeitos dos fármacos , Brânquias/metabolismo , Hepatopâncreas/efeitos dos fármacos , Hepatopâncreas/metabolismo , Concentração de Íons de Hidrogênio , Peroxidação de Lipídeos/efeitos dos fármacos , Toxinas Marinhas/metabolismo , Mytilus/genética , Mytilus/metabolismo , Temperatura
15.
Environ Sci Pollut Res Int ; 26(24): 24445-24461, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31228071

RESUMO

The study of the soil microbial community represents an important step in better understanding the environmental context. Therefore, biological characterisation and physicochemical integration are keys when defining contaminated sites. Fungi play a fundamental role in the soil, by providing and supporting ecological services for ecosystems and human wellbeing. In this research, 52 soil fungal taxa were isolated from in situ pilot reactors installed to a contaminated site in Czech Republic with a high concentration of hexachlorocyclohexane (HCH). Among the identified isolates, 12 strains were selected to evaluate their tolerance to different isomers of HCH by using specific indices (Rt:Rc; T.I.) and to test their potential in xenobiotic biotransformation. Most of the selected taxa was not significantly affected by exposure to HCH, underlining the elevated tolerance of all the tested fungal taxa, and different metabolic intermediates of HCH dechlorination were observed. The oxidative stress responses to HCH for two selected species, Penicillium simplicissimum and Trichoderma harzianum, were investigated in order to explore their toxic responses and to evaluate their potential functioning in bioremediation of contaminated environments. This research suggests that the isolated fungal species may provide opportunities for new eco-friendly, integrated and cost-effective solutions for environmental management and remediation, considering their efficient adaptation to stressful conditions.


Assuntos
Biotransformação , Fungos/metabolismo , Hexaclorocicloexano/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Biodegradação Ambiental , República Tcheca , Tolerância a Medicamentos , Ecossistema , Hexaclorocicloexano/análise , Isomerismo , Estresse Oxidativo , Penicillium/metabolismo , Solo , Poluentes do Solo/análise , Trichoderma/metabolismo
16.
J Neuropathol Exp Neurol ; 76(2): 135-150, 2017 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-28158670

RESUMO

Brain aging is accompanied by increased oxidative stress and what has been termed "neuroinflammation," which might contribute to age-related neurodegenerative diseases. We analyzed expression in the transcription of innate inflammatory response genes in eleven representative regions including frontal, parietal, inferior temporal, cingulate, occipital, entorhinal cortex, caudate, putamen, thalamus, substantia nigra, and cerebellar vermis in aging human brains. We probed members of the complement system, colony stimulating factor receptors, toll-like receptors, and pro- and anti-inflammatory cytokines in the brains of subjects with no neurological disease and neurofibrillary tangles (mean age: 47.1 ± 5.7 years) and those with no neurological disease and neurofibrillary pathology stages I-II (mean age: 70.6 ± 6.3 years). Although the entorhinal and frontal cortex were most altered, gene regulation patterns did not match regions with increased vulnerability. Analysis of false discovery rate thresholds revealed no differences for any gene in any region between the 2 groups, including cases in which individual comparisons analyzed using Student t or nonparametric tests showed apparent differences between groups. Moreover, gene expression of major anti-oxidative stress responses did not match neuroinflammation in aging or increased regional susceptibility to major neurodegenerative diseases.


Assuntos
Envelhecimento/genética , Envelhecimento/patologia , Inflamação/genética , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/patologia , Estresse Oxidativo/genética , Idoso , Encéfalo/patologia , Química Encefálica , Córtex Cerebral/metabolismo , Córtex Cerebral/patologia , Citocinas/biossíntese , Citocinas/genética , Feminino , Regulação da Expressão Gênica/genética , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes
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