• Animal Bioscience
• /
• v.36 no.7
• /
• pp.1022-1033
• /
• 2023

Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

• BMB Reports
• /
• v.56 no.4
• /
• pp.234-239
• /
• 2023

Thioredoxin-like protein 1 (TXNL1), one of the thioredoxin superfamily known as redox-regulator, plays an essential in maintaining cell survival via various antioxidant and anti-apoptotic mechanisms. It is well known that relationship between ischemia and oxidative stress, however, the role of TXNL1 protein in ischemic damage has not been fully investigated. In the present study, we aimed to determine the protective role of TXNL1 against on ischemic injury in vitro and in vivo using cell permeable Tat-TXNL1 fusion protein. Transduced Tat-TXNL1 inhibited ROS production and cell death in H₂O₂-exposed hippocampal neuronal (HT-22) cells and modulated MAPKs and Akt activation, and pro-apoptotic protein expression levels in the cells. In an ischemia animal model, Tat-TXNL1 markedly decreased hippocampal neuronal cell death and the activation of astrocytes and microglia. These findings indicate that cell permeable Tat-TXNL1 protects against oxidative stress in vitro and in vivo ischemic animal model. Therefore, we suggest Tat-TXNL1 can be a potential therapeutic protein for ischemic injury.

• Journal of Ginseng Research
• /
• v.47 no.3
• /
• pp.376-384
• /
• 2023

Background: Hepatic lipid disorder impaired mitochondrial homeostasis and intracellular redox balance, triggering development of non-alcohol fatty liver disease (NAFLD), while effective therapeutic approach remains inadequate. Ginsenosides Rc has been reported to maintain glucose balance in adipose tissue, while its role in regulating lipid metabolism remain vacant. Thus, we investigated the function and mechanism of ginsenosides Rc in defending high fat diet (HFD)-induced NAFLD. Methods: Mice primary hepatocytes (MPHs) challenged with oleic acid & palmitic acid were used to test the effects of ginsenosides Rc on intracellular lipid metabolism. RNAseq and molecular docking study were performed to explore potential targets of ginsenosides Rc in defending lipid deposition. Wild type and liver specific sirtuin 6 (SIRT6, 50721) deficient mice on HFD for 12 weeks were subjected to different dose of ginsenosides Rc to determine the function and detailed mechanism in vivo. Results: We identified ginsenosides Rc as a novel SIRT6 activator via increasing its expression and deacetylase activity. Ginsenosides Rc defends OA&PA-induced lipid deposition in MPHs and protects mice against HFD-induced metabolic disorder in dosage dependent manner. Ginsenosides Rc (20mg/kg) injection improved glucose intolerance, insulin resistance, oxidative stress and inflammation response in HFD mice. Ginsenosides Rc treatment accelerates peroxisome proliferator activated receptor alpha (PPAR-α, 19013)-mediated fatty acid oxidation in vivo and in vitro. Hepatic specific SIRT6 deletion abolished ginsenoside Rc-derived protective effects against HFD-induced NAFLD. Conclusion: Ginsenosides Rc protects mice against HFD-induced hepatosteatosis by improving PPAR-α-mediated fatty acid oxidation and antioxidant capacity in a SIRT6 dependent manner, and providing a promising strategy for NAFLD.

• Journal of The Korean Society of Integrative Medicine
• /
• v.11 no.4
• /
• pp.73-82
• /
• 2023

Purpose : Cymbopogon citratus, also known as lemongrass, has widely spread around the world and its essential oil is usually applied in food, perfume, and other industrial purposes. In addition, C. citratus has also been used for the treatment of inflammation, digestive disorders, and diabetes in traditional medicine. In this study, the antioxidative activity of C. citratus ethanol extract (CCEE) was analyzed in RAW 264.7 cells through the induction of one of phase II enzymes, heme oxygenase (HO)-1 by nuclear factor-erythroid 2 p45-related factor (Nrf)2, mitogen-activated protein kinase (MAPK), and phosphoinositide 3-kinase (PI3K)/Akt. Methods : The antioxidative activity of CCEE against oxidative stress and its underlying molecular mechanisms were analyzed by the cell viability assay, intracellular reactive oxygen species (ROS) formation assay, and Western blot analysis in RAW 264.7 cells. Results : The results exhibited that CCEE potently attenuated tert-butyl hydroperoxide (t-BHP) induced intracellular ROS levels in a dose-dependent manner without any cytotoxicity. CCEE treatment significantly induced the expression of HO-1 which is known for its antioxidative capacity. In addition, CCEE treatment significantly upregulated the expression of Nrf2, a corresponding transcription factor for the regulation of antioxidative enzymes, which was in accordance with the HO-1 overexpression. MAPK and PI3K/Akt were also evaluated for their important roles in the regulation of cellular redox homeostasis against oxidative damage. As a result, the potent HO-1 expression was mediated by not extracellular regulated kinase (ERK), c-Jun NH2 terminal kinase (JNK), p38, but phosphoinositide 3-kinase (PI3K) phosphorylation. To confirm the antioxidative activity of CCEE-induced HO-1 expression, oxidative damage was initiated by t-BHP and attenuated by CCEE treatment, which was identified by HO-1 selective inhibitor and inducer. Conclusion : Consequently, CCEE potently induced the HO-1-mediated antioxidative potential through the modulation of Nrf2 and PI3K/Akt signaling pathways in RAW 264.7 cells. These results suggest that CCEE could be a promising strategy for the mitigation against cellular oxidative damage.

• Economic and Environmental Geology
• /
• v.56 no.6
• /
• pp.745-764
• /
• 2023

In the Republic of Congo, Banded iron formations (BIFs) occur in two areas: the Chaillu Massif and the Ivindo Basement Complex, which are segments of the Archean Congo craton outcropping in the northwestern and southwestern parts of the country. They show interesting potential with significant mineral resources reaching 2 Bt and grades up to 60% Fe. BIFs consist mostly of oxide-rich facies (hematite/magnetite), but carbonate-rich facies are also highlighted. They are found across the country within the similar geological sequences composed of amphibolites, gneisses and greenschists. The Post-Archean Australian Shale (PAAS)-normalized patterns of BIFs show enrichment in elements such as SiO₂, Fe₂O₃, CaO, P₂O₅, Cr, Cu, Zn, Nb, Hf, U and depletion in TiO₂, Al₂O₃, MgO, Na₂O, K₂O, Sc, Th, Ba, Zr, Rb, Ni, V. REE diagrams show slight light REEs (rare earth elements; LREEs) compared to heavy REEs (HREEs), and positive La and Eu anomalies. The lithological associations, as well as the very high (Eu/Eu^*)_SN ratios> 1.8 shown by the BIFs, suggest that they are related to Algoma-type BIFs. The positive correlations between Zr and TiO₂, Al₂O₃, Hf suggest that the contamination comes mainly from felsic rocks, while the absence of correlations between MgO and Cr, Ni argues for negligeable contributions from mafic sources. Pr/Pr^* vs. Ce/Ce^* diagram indicates that the Congolese BIFs were formed in basins with redox heterogeneity, which varies from suboxic to anoxic and from oxic to anoxic conditions. They were formed through hydrothermal vents in the seawater, with relatively low proportions of detrital inputs derived from igneous sources through continental weathering. Some Congolese BIFs show high contents in Cr, Ni and Cu, which suggest that iron (Fe) and silicon (Si) have been leached through hydrothermal processes associated with submarine volcanism. We discussed their tectonic setting and depositional environment and proposed that they were deposited in extensional back-arc basins, which also recorded hydrothermal vent fluids.

• Journal of Plant Biotechnology
• /
• v.33 no.1
• /
• pp.1-9
• /
• 2006

Peroxiredoxins (Prxs) are ubiquitously distributed and play important functions in diverse cellular signaling systems. The proteins are largely classified into three groups, such as typical 2-Cys Prx, atypical 2-Cys Prx, and 1-Cys Prx, that are distinguished by their catalytic mechanisms and number of Cys residues. From the three classes of Prxs, the typical 2-Cys Prx containing the two-conserved Cys residues at its N-terminus and C-terminus catalyzes $H_2O_2$ with the use of thioredoxin (Trx) as an electron donor. During the catalytic cycle, the N-terminal Cys residue undergoes a peroxide-dependent oxidation to sulfenic acid, which can be further oxidized to sulfinic acid at the presence of high concentrations of $H_2O_2$ and a Trx system containing Trx, Trx reductase, and NADPH. The sulfinic acid form of 2-Cys Prx is reduced by the action of sulfiredoxin which requires ATP as an energy source. Under the strong oxidative or heat shock stress conditions, 2-Cys Prx in eukaryotes rapidly switches its protein structure from low-molecular-weight species to high-molecular-weight protein structures. In accordance with its structural changes, the protein concomitantly triggers functional switching from a peroxidase to a molecular chaperone, which can protect its substrate denaturation from external stress. In addition to its N-terminal active site, the C-terminal domain including 'YF-motif' of 2-Cys Prx plays a critical role in the structural changes. Therefore, the C-terminal truncated 2-Cys Prxs are not able to regulate their protein structures and highly resistant to $H_2O_2$-dependent hyperoxidation, suggesting that the reaction is guided by the peroxidatic Cys residue. Based on the results, it may be concluded that the peroxidatic Cys of 2-Cys Prx acts as an '$H_2O_2$-sensor' in the cells. The oxidative stress-dependent regulation of 2-Cys Prx provides a means of defense systems in cells to adapt stress conditions by activating intracellular defense signaling pathways. Particularly, 2-Cys Prxs in plants are localized in chloroplasts with a dynamic protein structure. The protein undergoes conformational changes again oxidative stress. Depending on a redox-potential of the chloroplasts, the plant 2-Cys Prx forms super-molecular weight protein structures, which attach to the thylakoid membranes in a reversible manner.

• Microbiology and Biotechnology Letters
• /
• v.34 no.2
• /
• pp.166-173
• /
• 2006

This research has been performed to clarify the relationship between hydrogeochemistry and bacterial community structure in groundwater contaminated with landfill leachate. We collected and analyzed samples from 5 sites such as leachate (KSG1-12), treated leachate (KSG1-16), two contaminated groundwaters (KSG1-07 and KSG1-08) and non-contaminated groundwater (KSG1-13). pH was 8.83, 8.04, 6.87, 6.87 and 6.53 in order; redox potential (Eh) 108, 202, 47, 200 and 154 mV; electric conductivity (EC) 3710, 894, 1223, 559 and 169.9 $\mu$S/cm; suspended solids (SS) 86.45, 13.74, 4.18, 0.24 and 11.91 mg/L. In KSG01-12, the ion concentrations were higher especially in $Cl^-$ and $HCO_3^-$ than other sites. The concentrations of Fe, Mn and $SO_4^{2-}$ were higher In KSG1-07 than in KSG1-08, and vise versa in $NO_3^{2-}$. In the comparison of DGGE fingerprint patterns, the similarity was highest between KSG1-13 and KSG1-16 (57.2%), probably due to common properties like low or none contaminant concentrations. Otherwise KSG1-08 showed lowest similarities with KSG1-13 (25.8%) and KSG1-12 (27.6%), maybe because of the degree of contamination. The most dominant bacterial species in each site were involved in $\alpha$-Proteobacteria (55.6%) in KSG1-12, $\gamma$-Proteobacteria (50.0%) in KSG1-16, $\beta$-Proteobacteria (66.7%) in KSG1-07, $\gamma$-Proteobacteria (54.5%) in KSG1-08 and $\beta$-Proteobacteria (36.4%) in KSG1-13. These results indicate that the microbial community structure might be changed according to the flow of leachate in grounderwater, implying changes in concentrations of pollutants, available electron accepters and/or other environmental conditions.

• Korean Journal of Environmental Agriculture
• /
• v.31 no.2
• /
• pp.104-112
• /
• 2012

BACKGROUND: Agricultural inputs (fertilizer and organic inputs) and water conditions can influence $CH_4$ and $CO_2$ emission from agricultural soils. This study was conducted to investigate the effects of agricultural inputs (fertilizer and organic inputs) under changing water regime on $CH_4$ and $CO_2$ emission from a soil in a laboratory incubation experiment. METHODS AND RESULTS: Four treatments were laid out: control without input and three type of agricultural inputs ($(NH_4)_2SO_4$, AS; pig manure compost, PMC; hairy vetch, HV). Fertilizer and organic inputs were mixed with 25 g of soil at 2.75 mg N/25 g soil (equivalent to 110 kg N/ha) in a bottle with septum, and incubated for 60 days. During the first 30-days incubation, the soil was waterlogged (1 cm of water depth) by adding distilled water weekly, and on 30 days of incubation, excess water was discarded then incubated up to 60 days without addition of water. Based on the redox potential, water regime could be classified into wetting (1 to 30 days), transition (31 to 40 days), and drying periods (41 to 60 days). Across the entire period, $CH_4$ and $CO_2$ flux ranged from 0 to 13.8 mg $CH_4$/m/day and from 0.4~1.9 g $CO_2$/m/day, and both were relatively higher in the early wetting period and the boundary between transition and drying periods. During the entire period, % loss of C relative to the initial was highest in HV (16.4%) followed by AS (8.1%), PMC (7.5%), and control (5.4%), indicating readily decomposability of HV. Accordingly, both $CH_4$ and $CO_2$ fluxes were greatest in HV treatment. Meanwhile, the lower $CH_4$ flux in AS and PMC treatments than the control was ascribed to reduction in $CH_4$ generation due to the presence of oxidized compounds such as ${SO_4}^{2-}$, $Fe^{3+}$, $Mn^{4+}$, and ${NO_3}^-$ that compete with precursors of $CH_4$ for electrons. CONCLUSION: Green manure such as HV can replace synthetic fertilizer in terms of N input, however, it may increase $CH_4$ emission from soils. Therefore, co-application of green manure and livestock manure compost needs to be considered in order to achieve satisfactory N supply and to mitigate $CH_4$ and $CO_2$ emission.

• Applied Biological Chemistry
• /
• v.45 no.2
• /
• pp.84-91
• /
• 2002

If contaminated river water is sprayed over the floodplain, organic matter and nitrogen would be removed by microbial processes in the rhizosphere of vegetation during the filtration through soil. In this study we tested the organic matter and nitrogen removal from contaminated river water by the floodplain filtration. Model system of floodplain was constructed using a PVC pipe (15 cm i.d. ${\times}$ 150 cm L) which was packed with a loamy sand soil collected from a floodplain in Nakdong river. The model system was instrumented with soil solution samplers and gas samplers. A river water collected from Omogcheon in Kyongsan was sprayed from top of the model system at three different rates. The concentration of organic matter, DO, $NO_3^-$, $NO_2^-$, $NH_4^+$, $N_2$ and $N_2O$, and redox potential were measured as a function of soil depth for 24 days after the system reached a steady state. When river water was sprayed at the rates of 40.8 and 68.0 $l/m^2/day$, a significant reductive condition for denitrification was developed at below 5-cm depth of the soil. When the water reached at 90-cm depth of the soil, COD and concentration of inorganic nitrogen were lowered, on an average, from 18.7 to 5 mg/l and from 2.7 to 0.4 mg/l, respectively. $N_2$ comprised most of the N gas evolved from denitrification and $N_2O$ concentrations emitted at the surface of soil were less than 1 {\mu}l/l. The effective removal of organic matter and nitrogen by the filtration in the model system of floodplain demonstrates that the native floodplains, which include rhizosphere of vegetation at the top soil, could be more effective in the treatment of contaminated river waters and other industrial waste waters containing high concentration of organic matter and nitrogen.

• Clean Technology
• /
• v.25 no.1
• /
• pp.46-55
• /
• 2019

CdS and CdZnS/ZnO materials were prepared using precipitation method and used as photocatalysts for the photocatalytic degradation of rhodamine B (RhB) under visible light irradiation. The prepared photocatalysts were also characterized by XRD and UV-vis DRS. The results indicated that the photocatalysts with intended crystalline structures were successfully obtained and both the CdS and CdZnS/ZnO can absorb visible light as well as UV. The photocatalytic activities were examined with the addition of scavenger for various active chemical species and the difference of reaction mechanisms over the catalysts were discussed. The $CH_3OH$, KI and p-benzoquinone were used as scavengers for ${\cdot}OH$ radical, photogenerated positive hole and ${\cdot}O_2{^-}$ radical, respectively. The CdS and CdZnS/ZnO showed different photocatalytic degradation mechanisms of RhB. It can be postulated that ${\cdot}O_2{^-}$ radical is the main active species for the reaction over CdS photocatalyst, while the photogenerated positive hole for CdZnS/ZnO photocatalyst. As a result, the predominant reaction pathways over CdS and CdZnS/ZnO photocatalysts were found to be the dealkylation of chromophore skeleton and the cleavage of the conjugated chromophore structure, respectively. The above results may be mainly ascribed to the difference of band edge potential of conduction and valence bands in CdS, CdZnS and ZnO semiconductors and the redox potentials for formation of active chemical species.