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At least 487 records · Page 27

Sub-Inhibitory Fosmidomycin Exposures Elicits Oxidative Stress in Salmonella enterica Serovar typhimurium LT2

Fosmidomycin is a time-dependent nanomolar inhibitor of methylerythritol phosphate (MEP) synthase, which is the enzyme that catalyzes the first committed step in the MEP pathway to isoprenoids. Importantly, fosmidomycin is one of only a few MEP pathway-specific inhibitors that exhibits antimicrobial activity. Most inhibitors identified to date only exhibit activity against isolated pathway enzymes. The MEP pathway is the sole route to isoprenoids in many bacteria, yet has no human homologs. The development of inhibitors of this pathway holds promise as novel antimicrobial agents. Similarly, analyses of the bacterial response toward MEP pathway inhibitors provides valuable information toward the understanding of how emergent resistance may ultimately develop to this class of antibiotics. We have examined the transcriptional response of Salmonella enterica serovar typhimurium LT2 to sub-inhibitory concentrations of fosmidomycin via cDNA microarray and RTPCR. Within the regulated genes identified by microarray were a number of genes encoding enzymes associated with the mediation of reactive oxygen species (ROS). Regulation of a panel of genes implicated in the response of cells to oxidative stress (including genes for catalases, superoxide dismutases, and alkylhydrogen peroxide reductases) was investigated and mild upregulation in some members was observed as a function of fosmidomycin exposure over time. The extent of regulation of these genes was similar to that observed for comparable exposures to kanamycin, but differed significantly from tetracycline. Furthermore, S. typhimurium exposed to sub-inhibitory concentrations of fosmidomycin displayed an increased sensitivity to exogenous H 2 O 2 relative to either untreated controls or kanamycin-treated cells. Our results suggest that endogenous oxidative stress is one consequence of exposures to fosmidomycin, likely through the temporal depletion of intracellular isoprenoids themselves, rather than other mechanisms that have been proposed to facilitate ROS accumulation in bacteria (e.g. cell death processes or the ability of the antibiotic to redox cycle).

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The Combined Effects of Arbuscular Mycorrhizal Fungi (AMF) and Lead (Pb) Stress on Pb Accumulation, Plant Growth Parameters, Photosynthesis, and Antioxidant Enzymes in Robinia pseudoacacia L.

Arbuscular mycorrhizal fungi (AMF) are considered as a potential biotechnological tool for improving phytostabilization efficiency and plant tolerance to heavy metal-contaminated soils. However, the mechanisms through which AMF help to alleviate metal toxicity in plants are still poorly understood. A greenhouse experiment was conducted to evaluate the effects of two AMF species (Funneliformis mosseae and Rhizophagus intraradices) on the growth, Pb accumulation, photosynthesis and antioxidant enzyme activities of a leguminous tree (Robinia pseudoacacia L.) at Pb addition levels of 0, 500, 1000 and 2000 mg kg -1 soil. AMF symbiosis decreased Pb concentrations in the leaves and promoted the accumulation of biomass as well as photosynthetic pigment contents. Mycorrhizal plants had higher gas exchange capacity, non-photochemistry efficiency, and photochemistry efficiency compared with non-mycorrhizal plants. The enzymatic activities of superoxide dismutase (SOD), ascorbate peroxidases (APX) and glutathione peroxidase (GPX) were enhanced, and hydrogen peroxide (H 2 O 2 ) and malondialdehyde (MDA) contents were reduced in mycorrhizal plants. These findings suggested that AMF symbiosis could protect plants by alleviating cellular oxidative damage in response to Pb stress. Furthermore, mycorrhizal dependency on plants increased with increasing Pb stress levels, indicating that AMF inoculation likely played a more important role in plant Pb tolerance in heavily contaminated soils. Overall, both F. mosseae and R. intraradices were able to maintain efficient symbiosis with R. pseudoacacia in Pb polluted soils. AMF symbiosis can improve photosynthesis and reactive oxygen species (ROS) scavenging capabilities and decrease Pb concentrations in leaves to alleviate Pb toxicity in R. pseudoacacia. In conclusion, our results suggest that the application of the two AMF species associated with R. pseudoacacia could be a promising strategy for enhancing the phytostabilization efficiency of Pb contaminated soils.

59 BASIC BIOLOGICAL SCIENCES↗

Antioxidant metabolism variation associated with alkali-salt tolerance in thirty switchgrass (Panicum virgatum) lines

Soil salinization is a major factor limiting crop growth and development in many areas. Switchgrass (Panicum virgatum L.) is an important warm-season grass species used for biofuel production. The objective of this study was to investigate antioxidant metabolism, proline,and protein variation associated with alkali-salt tolerance among 30 switchgrass lines and identify metabolic markers for evaluating alkali-salt tolerance of switchgrass lines. The grass lines were transplanted into plastic pots containing fine sand. When the plants reached E5 developmental stage, they were subjected to either alkali-salt stress treatment (150 mM Na + and pH of 9.5) or control (no alkali-salt stress) for 20 d. The 30 switchgrass lines differed in alkali-salt tolerance as determined by the level of leaf malondialdehyde (MDA), antioxidant enzyme activity [(superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX)], proline and protein. Alkali-salt stress increased MDA, proline, SOD, reduced CAT activity, but its effect on protein and APX varied depending on lines. Wide variations in the five parameters existed among the 30 lines. In general, the lines with higher CAT activity and lower proline content under alkali-salt stress had less MDA, exhibiting better alkali-salt tolerance. Among the five parameters, CAT can be considered as valuable metabolic markers for assessment of switchgrass tolerance to alkali-salt stress.

09 BIOMASS FUELS↗

In vitro toxicity assessment of uranium particulates on different human lung epithelial cell models

Inhalation of uranium aerosols produced via human activities such as mining can pose a threat to human respiratory systems. Uranium oxide particulates emit short-range alpha particles that elicit DNA and direct damage, beyond associated physiochemical heavy-metal toxicity, to internal epithelial tissues. The availability of reliable in vitro models to study radiation exposure can greatly enhance our ability to understand and combat the biological impacts of exposure. However, the toxicological effects of alpha emissions and/or the oxidation states of uranium particulates vary across different human lung epithelial cell models and have not been systematically compared. We have endeavored to address this limitation by comparing impacts in three different human lung cell models: primary human bronchial and tracheal epithelial cells, primary human small airway epithelial cells, and human adenocarcinoma alveolar basal epithelial cells. Other studies have mainly investigated the toxicity of depleted uranium. Here, we compared the exposure of uranium oxide particulates (U 3 O 8 and UO 3 ) of different enrichment states on the chosen cell systems. Each cell model was exposed to 0.1, 1, 10, 50, 100, and 500 µg/mL of depleted U 3 O 8 , highly-enriched U 3 O 8 , and natural UO 3 particulates for 24 hours in submerged monolayer cultures. We compared viability and superoxide dismutase activity results across cell lines and uranium enrichment/ oxidative states. The results showed that 1) the oxide state of the particulates affected cell viability, implying that uranium’s different oxidation states contribute to different toxicological responses, and 2) each cell model reacts differently when exposed to uranium oxides, which may provide insights into the mechanistic processes associated with the exposure of radiological particulates on different biological systems. For instance, increased uranium enrichment corresponds to increased toxicity for the primary cells, but not for the immortalized cells. Our study shows that a holistic approach that incorporates similarities between model systems and types of radionuclides is required to truly develop empirical solutions for radiation exposure.

59 BASIC BIOLOGICAL SCIENCES↗

Importance of Microbial Activity On Groundwater Iodate and Organo-Iodine Speciation and Mobility At Two DOE Sites

Iodine (I) occurs in multiple oxidation states in aquatic systems in the form of organic and inorganic species (iodide and iodate). This fact leads to complex biogeochemical cycling of Iodine and its long-lived isotope, 129 I, a major by-product of nuclear fission. Results from our newly developed, sensitive and rapid method for speciated isotopic ratios ( 129 I/ 127 I) via GC-MS, which compare favorably with Accelerator Mass Spectroscopy, demonstrate that the mobility of 129 I species greatly depends on the type of I species and its concentration, pH, and sediment redox state. At ambient concentrations (~10 7 M), I- and IO -3 are significantly retarded by sorption to mineral surfaces and covalent binding to aromatic moieties in natural organic matter (NOM), even when NOM is present at low concentrations such as occur at Hanford. At concentrations traditionally examined in sorption studies (≥ 10 -4 M), I- travels along with the water. Iodate removal can also occur through incorporation into CaCO 3 crystal lattice, e.g., at the Hanford Site. Removal of iodine from the groundwater through interaction with NOM is complicated by the release of mobile organo-I species, as was observed at SRS and Hanford. A small fraction of NOM that is bound to iodine can behave as a mobile organo-I source, a process that we were able to numerically simulate. Field and laboratory studies evaluating the cause for steady increases in 129 I concentrations (up to 1000 pCi L-1) emanating from radiological basins at SRS indicate that an increase of 0.7 pH units in groundwater over 17 years can account for the observed increased groundwater 129 I concentrations. Bacteria from a 129 I-contaminated aerobic aquifer at the F-area of SRS can accumulate I- at environmentally relevant concentrations (10 -7 M), and enzymatically oxidize I-, which together with microbially produced MnO 2 and superoxide or organic acids can significantly contribute to organo-iodine formation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Final report for project "Effects of Low-Dose Irradiation on NFkB Signaling Networks and Mitochondria"

Low dose ionizing radiation effects are difficult to study in human population because of the numerous confounding factors such as genetic and lifestyle differences. Research in mammalian model systems and in vitro is generally used in order to overcome this difficulty. In this program project three projects have joined together to investigate effects of low doses of ionizing radiation. These are doses at and below 10 cGy of low linear energy transfer ionizing radiation such as X-ray and gamma rays. This project was focused on cellular signaling associated with nuclear factor kappa B (NFkB) and mitochondria - subcellular organelles critical for cell aging and aging-like changes induced by ionizing radiation. In addition to cells in culture this project utilized animal tissues accumulated in a radiation biology tissue archive housed at Northwestern University (http://janus.northwestern.edu/janus2/index.php). Major trust of Project 1 was to gather all of the DoE sponsored irradiated animal (mouse, rat and dog) data and tissues under one roof and investigate mitochondrial DNA changes and micro RNA changes in these samples. Through comparison of different samples we were trying to delineate mitochondrial DNA quantity alterations and micro RNA expression differences associated with different doses and dose rates of radiation. Historic animal irradiation experiments sponsored by DoE were done in several national laboratories and universities between 1950’s and 1990’s; while these experiments were closed data and tissues were released to Project 1. Project 2 used cells in culture to investigate effects that low doses or radiation have on NFκB and its target genes manganese superoxide dismutase (MnSOD) and genes involved in cell cycle: Cyclins (B1 and D1) and cyclin dependent kinases (CDKs). Project 3 used cells in culture such as “normal” human cells (breast epithelial cell line MCF10A cells and skin keratinocyte cells HK18) and mouse embryo fibroblast (mef) cells to focus on role of NFkB protein and several other proteins such as survivin (BIRC5) in radiation dependent regulation of tumor necrosis factor alpha (TNFα) and its downstream signaling.

59 BASIC BIOLOGICAL SCIENCES↗

Final Report/Liox Power Inc.

The rechargeable lithium-oxygen battery has attracted attention due to its large theoretical energy density compared to modern lithium-ion batteries. This large energy density is attributed to the reaction of lithium with molecular oxygen to form lithium peroxide, which grows on the surface of the cathode. While this is a promising chemistry, there are many practical challenges that remain to be solved, such as the decomposition of organic electrolyte in the presence of superoxide anions and large overpotentials on charge.

25 ENERGY STORAGE↗

Life on the Edge: Microbes in Rock Varnish

The newly discovered high concentrations of manganese on Mars open up new possibilities for habitability on that planet [1, 2]. On Earth, there is a close association between Mn deposits and the presence of Mn- and Fe- oxidizing microbes [3-7]; as such, Mn is considered a principal biosignature for Mars [8]. The most common terrestrial Mn-rich surface material on Earth is desert varnish, a dark, shiny coating on rocks in arid locations. Microorganisms occupy various niches within rock varnish, and it is probable that the concentration of Mn in many rock varnishes is mediated by microbial activity [6, 9, 10]; however, the relationship between microbes and varnish remains a source of long-standing controversy [11]. Do microorganisms drive the formation of varnish, oxidizing Mn and Fe to gain energy, or do they instead utilize varnish as a habitat? This long-standing question has important implications for our understanding of how life on Earth has evolved to capture and harness energy from the physical environment. Additionally, it will greatly influence the search on Mars for the signatures of life that may be present in rock varnish cannot be definitively identified on Mars without first being identified on Earth. Here we explored the microbial species and processes involved in the habitation of rock varnish and identified organic biosignatures that, in concert with trace element and mineralogy, could be used to distinguish the biogenic and abiogenic origins of terrestrial Mn-rich surfaces so that we may then apply the knowledge gained in this work to current Martian datasets from ChemCam. Each of these goals has important implications for our understanding of how life on Earth has evolved to capture and harness energy from the physical environment. A combinatorial experimental approach, utilizing microscopy, high-throughput DNA sequencing, culturing and physiological assays, were used to characterize the microbial communities inhabiting varnished rocks across the Western U.S. Microbial communities inhabiting varnish were shaped by both location (larger contribution) and rock type. However, we found that the unique varnish environment selects for a core group of radiation- and desiccation-tolerant microorganisms across landscapes and rock types. The bacteria comprising this core group include a who’s who of radiation resistant organisms including Rubrobacter spp., Deinococcus spp., Hymenobaceter spp., and Chroococcidiopsis spp. Among the fungi, potential lichen-forming members of the Lecanoromycetes and rock-inhabiting Dothideomycetes were common. Importantly, we found that two of the survival mechanisms employed by these microorganisms are likely key drivers of Mn oxide formation and the biogenesis of varnish. First, we showed that the key cyanobacteria inhabiting the varnish accumulate copious amounts of low molecular weight Mn2+-conjugates, that are critical for their ability to withstand the toxic effects of reactive oxygen species generated upon radiation exposure. Secondly, the microbial community inhabiting varnished rocks were found to be particularly adept at producing siderophores, which are high-affinity chelating compounds secreted by bacteria and fungi that serve to sequester Fe2+ (and Mn2+) from oxidizing environments for transport into cells as essential micronutrients for growth and survival. When tightly bound to organic ligands, Mn2+ is susceptible to oxidation to Mn3+-ligand under atmospheric oxygen and at slightly basic pH levels. Further, Mn3+-ligand can be oxidized to MnO2 (manganese oxide) when it comes into contact with reactive oxygen species (e.g., superoxide radical) formed during the photolysis of water during periods of high UV exposure. The data points towards a scenario whereby cyanobacterial biofilms, and associated radiation resistant, heterotrophic bacteria, comprise the central hub of the varnish community. These communities depend on light and water to slowly establish on the rock surface where they accumulate and concentrate Mn for survival. When the biofilms dry out, mineral rinds rich in Mn oxides form which serve as a habitat for the successive rounds of microbial growth and Mn oxide formation. Copious production and excretion of siderophores, which are extremely stable in the environment, coats the rock surface with a longlived catalyst for Mn oxidation. By discovering these two completely novel mechanisms for biogenic varnish formation, we have gained the underlying knowledge necessary to develop a suite of organic and inorganic biosignatures to distinguish biogenic and abiogenic origins of Mnrich surfaces on Earth or Mars.

59 BASIC BIOLOGICAL SCIENCES↗

Combined Experimental and Computational Efforts to Establish Ion Mobility, Solubility and Stability of Functional Liquids for Electrochemical Energy Storage

This work provides a computation-driven investigation of the stability of organic electrolytes for lithium-air batteries. Electrolyte instability is currently a key challenge that limits practical use of aprotic Li-air batteries, and the chemical processes that cause this instability are often kinetically-driven. Computational screening for kinetic stability involves the determination of reaction barriers for the numerous potential reaction mechanisms, barriers that are challenging to calculate due to the difficulty of locating transition state structures. Here we screen a broad set of substituted electrolytes for susceptibility to nucleophilic attack by superoxide. We find that carbonates are not typically expected to be stable and that sulfones are generally stable, validating literature trends. We study the effects of chemical functionalization with electron-donating and withdrawing groups and their interplay with steric factors, identifying functional groups and other chemical modifications that increase stability in these groups. User-input driven transition state identification is used for these initial calculations, and an automated computational pipeline is subsequently presented and validated as a means to perform further high-throughput searches across mechanisms and chemistries. The pipeline integrates cheminformatics-based reaction encoding, relaxed potential energy scans, and nudged elastic band calculations for an end-to-end approach to barrier calculations. We review this automated search approach and its current limitations, and discuss challenges and further work.

25 ENERGY STORAGE↗

The Effects of Aging on Indices of Oxidative Stress and Apoptosis in the Female Fischer 344/Nnia X Brown Norway/BiNia Rat Heart

Oxidative-nitrosative stress may play a role in age-associated cardiovascular disease as implied by recent studies.However, limited research has been conducted using aged female rodent models. In this study, we examined hearts obtained from 6-, 26-, and 30-month old female Fischer 344/Nnia x Brown Norway/BiNia (F344xBN) rats in order to examine how aging affects levels of cardiac oxidative-nitrosative stress and apoptosis. Oxidative (superoxide anion and 4-HNE) and nitrosative (protein nitrosylation) stress markers were increased 180 ± 17 %, 110 ± 3 %, and 14 ± 2 %, respectively in 30-month hearts compared to the hearts of 6-month female rats. Coincident with these changes in oxidative-nitrosative stress, aging was also found to be associated with increases in the number of Tdt-mediated dUTP nick labeling (TUNEL)-positive cardiomyocytes, alterations in the Bax/Bcl-2 ratio, and elevated cleavage of caspase-3. Regression analysis demonstrates significant correlation in the age-associated changes markers of oxidative–nitrosative stress with changes in apoptotic signaling. The findings from this descriptive study imply that age-associated increases in mitochondrial-mediated apoptosis may be associated with the increase in oxidative-nitrosative stress in the aging F344xBN female heart.

60 APPLIED LIFE SCIENCES↗

Iron-mediated oxidation of methoxyhydroquinone at Rifle, Colorado

In this study, the oxidation of 2-methoxyhydroquinone (MH2Q) by ferric iron (Fe(III)) under dark conditions in the absence and presence of oxygen was investigated within a pH range of 4 – 6. While Fe(III) was capable of stoichiometrically oxidizing MH2Q under anaerobic conditions, catalytic oxidation of MH2Q was observed in the presence of O2 due to further cycling between oxygen, semiquinone radicals, and iron species. A detailed kinetic model was developed to describe the predominant mechanisms, which indicated that both the undissociated and mono-dissociated anions of MH2Q were kinetically active species towards Fe(III) reduction, with the mono-dissociated anion being the key species accounting for the pH dependence of the oxidation. The generated radical intermediates, namely semiquinone and superoxide, are of great importance in reaction chain propagation. The kinetic model may provide critical insight into the underlying mechanisms of the thermodynamic and kinetic characteristics of metal-organic interactions and assist in understanding and predicting the factors controlling iron and organic matter transformation and bioavailability in aquatic systems.

54 ENVIRONMENTAL SCIENCES↗

Streptococcus mutans Displays Altered Stress Responses While Enhancing Biofilm Formation by Lactobacillus casei in Mixed-Species Consortium

Like Streptococcus mutans , lactobacilli are commonly isolated from carious sites, although their exact role in caries development remains unclear. This study used mixed-species models to analyze biofilm formation by major groups of oral lactobacilli, including L. casei, L. fermentum, L. rhamnosus, L. salivarius ssp. salivarius, and L. gasseri . The results showed that lactobacilli did not form good biofilms when grown alone, although differences existed between different species. When grown together with S. mutans , biofilm formation by L. gasseri and L. rhamnosus was increased by 2-log ( P < 0.001), while biofilms by L. fermentum reduced by >1-log ( P < 0.001). L. casei enhanced biofilm formation by ~2-log when grown with S. mutans wild-type, but no such effects were observed with S. mutans deficient of glucosyltransferase GtfB and adhesin P1. Both S. mutans and L. casei in dual-species enhanced resistance to acid killing with increases of survival rate by >1-log ( P < 0.001), but drastically reduced the survival rates following exposure to hydrogen peroxide ( P < 0.001), as compared to the respective mono-species cultures. When analyzed by RNA-seq, more than 134 genes were identified in S. mutans in dual-species with L. casei as either up- or down-regulated when compared to those grown alone. The up-regulated genes include those for superoxide dismutase, NADH oxidase, and members of the mutanobactin biosynthesis cluster. Among the down-regulated genes were those for GtfB and alternative sigma factor SigX. These results further suggest that interactions between S. mutans and oral lactobacilli are species-specific and may have significant impact on cariogenic potential of the community.

Immunology↗

Quantitative measurements of reactive oxygen species partitioning in electron transfer flavoenzyme magnetic field sensing

Biological magnetic field sensing that gives rise to physiological responses is of considerable importance in quantum biology. The radical pair mechanism (RPM) is a fundamental quantum process that can explain some of the observed biological magnetic effects. In magnetically sensitive radical pair (RP) reactions, coherent spin dynamics between singlet and triplet pairs are modulated by weak magnetic fields. The resulting singlet and triplet reaction products lead to distinct biological signaling channels and cellular outcomes. A prevalent RP in biology is between flavin semiquinone and superoxide (O 2 •–) in the biological activation of molecular oxygen. This RP can result in a partitioning of reactive oxygen species (ROS) products to form either O 2 •– or hydrogen peroxide (H 2 O 2 ). Here, we examine magnetic sensing of recombinant human electron transfer flavoenzyme (ETF) reoxidation by selectively measuring O 2 •– and H 2 O 2 product distributions. ROS partitioning was observed between two static magnetic fields at 20 nT and 50 μT, with a 13% decrease in H 2 O 2 singlet products and a 10% increase in O 2 •– triplet products relative to 50 µT. RPM product yields were calculated for a realistic flavin/superoxide RP across the range of static magnetic fields, in agreement with experimental results. For a triplet born RP, the RPM also predicts about three times more O 2 •– than H 2 O 2 , with experimental results exhibiting about four time more O 2 •– produced by ETF. The method presented here illustrates the potential of a novel magnetic flavoprotein biological sensor that is directly linked to mitochondria bioenergetics and can be used as a target to study cell physiology.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Editorial: Rising stars in plant ROS/redox biology under abiotic stress conditions

Plants are constantly subjected to a wide range of abiotic stresses including salinity, high temperatures, drought, or cold, among others, that negatively impact their growth and development and cause crop yield losses worldwide (Lesk et al., 2016; Bailey-Serres et al., 2019). These stress conditions induce the accumulation of reactive oxygen species (ROS) such as hydrogen peroxide (H 2 O 2 ), superoxide ($O_{2•}$$^—$), singlet oxygen ($^1O_2$), or hydroxyl radical (HO·). ROS play a key role in stress sensing and signaling, as well as in the activation of different mechanism involved in plant acclimation to different stresses (Mittler et al., 2022). However, ROS levels that are too high are cytotoxic (Mittler, 2017). To prevent cellular oxidation by ROS, an interplay between ROS production, scavenging and transport maintains ROS at controlled concentrations, as well as drives ROS signaling responses involved in plant acclimation to stress (Baxter et al., 2014; Mittler et al., 2022).

protein carbonylation↗

Dietary Strategies to Modulate the Health Condition and Immune Responses in Gilthead Seabream (Sparus aurata) Juveniles Following Intestinal Inflammation

Several feed additives have proved to be beneficial in eliciting fish health. Β-glucans and curcumin are compounds with immunomodulatory capacities known to increase growth performance, stimulate immunity, improve general health, and enhance disease resistance in fish. The present study aimed to evaluate the effects of dietary Phaeodactylum tricornutum extracts rich in β-glucans and curcumin on gilthead seabream health status prior to and following an intestinal inflammatory stimulus. Three experimental diets were formulated: a practical commercial-type diet (CTRL), a CTRL diet supplemented with 1% microalgae-derived β-glucans extract (BG), and a CTRL diet supplemented with 0.2% of curcumin (CUR). After 30 days of the feeding trial, fish were sampled and subjected to an oral administration of 1% dextran sodium sulphate (DSS) to induce intestinal inflammation. Four groups were considered: a group of fish continued to be fed on the CTRL diet while the remaining groups were exposed to DSS, including CTRL-D (CTRL + DSS), BG-D (BG + DSS), and CUR-D (CUR + DSS), for 6 days. Growth, plasma and gut humoral immunity, liver and gut oxidative stress biomarkers, and intestinal gene expression were evaluated. No significant differences were found in growth after 30 days of feeding; however, seabream fed BG had decreased anti-protease activity and nitric oxide concentration in plasma while those fed CUR had increased mRNA levels of the tnfα, csf1r, and hep genes compared to those fed CTRL. After the inflammatory stimulus, hematocrit was enhanced in fish fed BG-D and CUR-D while red blood cell counts increased in those fed CTRL-D. Superoxide dismutase activity decreased in the intestine of all DSS groups while lipid peroxidation increased in the gut of fish fed CTRL-D and BG-D compared to CTRL. Moreover, the mRNA expression levels of csfr1 and sod decreased in fish fed CTRL-D and BG-D compared to CTRL, respectively. Despite the mild intestinal inflammatory condition induced by DSS, CUR was able to partially ameliorate its effects, improving the hematological profile and assisting against the oxidative stress.

Teixeira, Carla↗

Supplemental Microalgal DHA and Astaxanthin Affect Astaxanthin Metabolism and Redox Status of Juvenile Rainbow Trout

Microalgal docosahexaenoic acid (DHA) and astaxanthin (AST) may substitute for fish oil and synthetic AST in aquafeeds. This study explored the effects and mechanisms of those substitutions on AST metabolism and redox status of rainbow trout fed plant protein meal (PM)- or fishmeal (FM)-based diets. Two parallel experiments (PM vs. FM) were performed with 612 juvenile rainbow trout for 16 weeks as a 2 × 3 factorial arrangement of treatments with two AST sources (synthetic (SA) vs. microalgal (AA), at 80 mg/kg) and three levels (0, 50, and 100%) of fish oil substitutions with DHA-rich microalgae. The fish oil substitutions exhibit main effects (p < 0.05) and/or interactive effects (p < 0.05) with the source of AST on AST deposition, malondialdehyde and glutathione concentrations, and mRNA levels and activities of major redox enzymes (glutathione reductase (GR), glutathione peroxidase (GPX), glutathione S-transferase (GST), and superoxide dismutase (SOD)) in the muscle and liver of trout fed both diet series. The AST source produced only differences in tissue AST deposition (p < 0.05) and number of metabolites. In conclusion, the substitutions of fish oil by the DHA-rich microalgae exerted more impacts than those of SA by AA on redox status and functional expression of antioxidant enzymes in the tissues of rainbow trout.

59 BASIC BIOLOGICAL SCIENCES↗

Detection Technologies for Reactive Oxygen Species: Fluorescence and Electrochemical Methods and Their Applications

Reactive oxygen species (ROS) have been found in plants, mammals, and natural environmental processes. The presence of ROS in mammals has been linked to the development of severe diseases, such as diabetes, cancer, tumors, and several neurodegenerative conditions. The most common ROS involved in human health are superoxide (O 2 •-), hydrogen peroxide (H 2 O 2 ), and hydroxyl radicals (•OH). Organic and inorganic molecules have been integrated with various methods to detect and monitor ROS for understanding the effect of their presence and concentration on diseases caused by oxidative stress. Among several techniques, fluorescence and electrochemical methods have been recently developed and employed for the detection of ROS. This literature review intends to critically discuss the development of these techniques to date, as well as their application for in vitro and in vivo ROS detection regarding free-radical-related diseases. Moreover, important insights into and further steps for using fluorescence and electrochemical methods in the detection of ROS are presented.

59 BASIC BIOLOGICAL SCIENCES↗

De novo Characterization of the Platycladus orientalis Transcriptome and Analysis of Photosynthesis-Related Genes during Aging

In China, Platycladus orientalis has a lifespan of thousands of years. The long lifespan of these trees may be relevant for the characterization of plant aging at the molecular level. However, the molecular mechanism of the aging process of P. orientalis is still unknown. To explore the relationship between age and growth of P. orientalis , we analyzed physiological changes during P. orientalis senescence. The malondialdehyde content was greater in 200-, 700-, and 1100-year-old ancient trees than in 20-year-old trees, whereas the peroxidase and superoxide dismutase activities, as well as the soluble protein content, exhibited the opposite trend. Furthermore, we performed a de novo transcriptome assembly using RNA-Seq and obtained 48,044 unigenes with an average length of 896 bp. A total of 418 differentially expressed genes were identified in different stages of aging of P. orientalis . Clustering analysis revealed distinct timepoints at which the oxidation–reduction and photosynthesis pathways changed. Eight clusters with distinct expression patterns were identified. The expression levels of photosynthesis-, oxidation–reduction-, and transporter-related genes were down-regulated, whereas those of transcription-, signaling-, and senescence-related genes were up-regulated during aging. In addition, consistent with the most obviously down-regulated genes of photosynthesis-related genes, the photosynthetic indexes including chlorophyll a and b levels decreased steadily during P. orientalis aging. This study combined transcriptome with physiological and biochemical data, revealing potential candidate genes influencing senescence during P. orientalis aging.

59 BASIC BIOLOGICAL SCIENCES↗