• Korean Journal of Plant Resources
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• v.29 no.2
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• pp.163-170
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• 2016

Antioxidant activity and inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Cone of Red Pine (Pinus densiflora) were investigated to find utilization of Cone, by-product of Red Pine, thrown out after berry shatter, as a new natural plant resource. Cone from P. densiflora was extracted with methanol (MeOH) and separated to petroleum ether, ethyl acetate and water fraction. Among them, ethyl acetate fraction was used. The antioxidant activity was conducted by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay, Fe²⁺ chelating assay and reducing power assay. The inhibitory effect on oxidative DNA damage was determined by DNA cleavage assay using φX-174 RF I plasmid. The results of DPPH and ABTS radical scavenging activity at 200 ㎍/㎖ of extracts were 86.50% and 95.80% respectively, which were similar figures compared with L-ascorbic acid as control. Fe²⁺ chelating activity was 77.96% and reducing power was 0.77 at 200 ㎍/㎖. Total phenolic component was 27.29±0.3 ㎎/g and Vitamin C content was 1.84±0.1 ㎎/g. Also ethyl acetate fraction from Cone has inhibitory effect, using φX-174 RF I plasmid on DNA cleavage assay. In conclusion, Cone, by-product of P. densiflora, showed high antioxidant activity and inhibitory effect on oxidative DNA damage. Therefore this study suggests Cone, useless by-product, can be developed as a new natural plant resource with lots of utilization such as an effective antioxidant, natural medicine, food, cosmetics and so on.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.131-137
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• 2024

Background: Porcine embryonic development is widely utilized in the medical industry. However, the blastocyst development rate in vitro is lower compared to in vivo. To address this issue, various supplements are employed. Extracellular vesicles (EVs) play the role of communicators that carry many bioactive cargoes. Additionally, the contents of EVs can vary on the estrous cycle. Methods: We compared the effects of adding EVs derived from porcine uterine fluid (UF), categorized as non-EV (G1), EVs in estrus (G2) and EVs in diestrus (G3). After in vitro culture (IVC) was performed in three different groups, cleavage rate and blastocyst development rate were examined. In addition, glutathione (GSH) and reactive oxygen species (ROS) levels were measured 2 days after activation to assess oxidative stress. Results: Using NTA and cryo-TEM, we confirmed the presence of EVs with sizes ranging from 30 nm to 200 nm, that the particles were suitable for analysis for analysis. In IVC data, the highest cleavage rate was observed in G2, which was significantly different from G1 but not significantly different from the next highest, G3. Similarly, the highest blastocyst development rate was observed in G2, which was significantly different from G1 but not significantly different from the next highest, G3. Conclusions: These results indicate that estrus derived EVs contain biofactors beneficial for early blastocyst development, including GSH which protects the blastocyst from oxidative stress. Additionally, although diestrus-derived EVs are expected to have some effect on blastocyst development, it appeared to be less effective than estrus-derived EVs.

• Journal of Microbiology
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• v.38 no.2
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• pp.53-61
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• 2000

Hydroxybenzoic acids are the most important intermediates in the degradative pathways of various aromatic compounds. Microorganisms catabolize aromatic compounds by converting them to hydroxylated intermediates and then cleave the benzene nucleus with ring dioxygenases. Hydroxylation of the benzene nucleus of an aromatic compound is an essential step for the initiation and subsequent disintegration of the benzene ring. The incorporation of two hydroxyl groups is essential for the labilization of the benzene nucleus. Monohydroxybenzoic acids such as 2-hydroxybenzoic acid, 3-hydroxybenzoic acid, and 4-hydrosybenzoic acid, opr pyrocattechuic acid that are susceptible for subsequent oxygenative cleavage of the benzene ring. These terminal aromatic intermediates are further degraded to cellular components through ortho-and/or meta-cleavage pathways and finally lead to the formation of constituents of the TCA cycle. Many groups of microorganisms have been isolated as degraders of hydroxybenzoic acids with diverse drgradative routes and specific enzymes involved in their metabolic pahtway. Various microorganisms carry out unusual non-oxidative decarboxylation of aromatic acids and convert them to respective phenols which have been documented. Futher, Pseudomonas and Bacillus spp. are the most ubiquitous microorganisms, being the principal components of microflora of most soil and water enviroments.

• Bulletin of the Korean Chemical Society
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• v.21 no.9
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• pp.923-928
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• 2000

[FeIII(BBA)DBC]ClO4 as a new functional model for catechol dioxygenases has been synthesized, where BBA is a bis(benzimidazolyl-2-methyl)amine and DBC is a 3,5-di-tert-butylcatecholate dianion.The BBA complex has a structuralfeature that iron cent er has a five-coordinate geometry similar to that of catechol dioxygenase-substrate complex.The BBA complex exhibits strong absorptionbands at 560 and 820 nm in CH3CN which are assigned to catecholate to Fe(III) charge transfer transitions. It also exhibits EPR signals at g = 9.3 and 4.3 which are typical values for the high-spin FeIII (S = 5/2) complex with rhombicsymmetry. Interestingly, the BBA complex reacts with O2 within an hour to afford intradiol cleavage (35%) and extradiol cleavage (60%) products. Surprisingly, a green color intermediate is observed during the oxygenation process of the BBA com-plex in CH3CN. This green intermediate shows a broad isotropic EPR signal at g = 2.0. Based on the variable temperature EPR study, this isotropic signalmight be originated from the [Fe(III)-peroxo-catecholate] species havinglow-spin FeIII center, not from the simple organic radical. Consequently,it allows O2 to bind to iron cen-ter forming the Fe(III)-superoxide species that converts to the Fe(III)-peroxide intermediate. These present data can lead us tosuggest that the oxygen activation mechanism take place for the oxidative cleavingcatechols of the five-coordinate model systems for catechol dioxygenases.

• Journal of the Korean Society of Food Culture
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• v.35 no.4
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• pp.400-405
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• 2020

Evodiae Fructus is the dried unripe fruit of Evodia rutaecarpa, and has traditionally been used for treating stomachache and diarrhea. Evodiamine and rutaecarpine, the major biologically active compounds of Evodiae Fructus, are reported to have anti-oxidative and anti-inflammatory effects, as well as inhibit proliferation and metastasis of various cancer cells. The current study investigates the anti-oxidative and anti-cancer effects of the Evodiae Fructus extract, considering varying concentrations of methanol extraction (40, 80, and 95%). High contents of total phenolic compounds were determined in the order of extracts 80, 95, and 40%. Evaluating contents of the 95, 80, and 40% extracts revealed 36.77, 7.29, and 1.86 ㎍/mg evodiamine, respectively, and 53.02, 17.16, and 3.79 ㎍/mg rutaecarpine, respectively, with the highest content of both compounds obtained in the 95% extract. DPPH radical scavenging activity was observed to be inversely proportional to the contents of total phenolic compounds, with decreasing SC₅₀ values obtained in the order 80, 95, and 40% extract. The 95 and 80% extracts exerted toxicity to AGS gastric cancer cells, but the 40% extract was non-toxic. Evodiamine is a known anti-cancer agent, and could be responsible for the observed toxicity. Cleavage of PARP, and Caspase-3, -7, -8 and -9 was observed in the 95% extract-treated AGS cells, indicating that cell toxicity exerted by the 95% extract could be attributed to apoptosis.

• Herbal Formula Science
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• v.21 no.2
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• pp.63-71
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• 2013

Objectives : Artemisia princeps is used as moxa in moxibustion and traditional herbal medicine. And its extracts or compounds is known to have an efficacy of antioxidant, anti-diabete, anti-cancer, anti-inflammation and neuroprotection. This study was performed to investigate the cytoprotective effect of Artemisia princeps extract (APE) against arachidonic acid (AA)+iron-induced oxidative stress on HepG2 cell. Methods : The effects of APE on cell viability has been assessed using MTT assay. And flow cytometric analysis was performed to estimate APE's effects on mitochondrial function. To investigate its underlying mechanism, related protein was analysed by using immunoblot analysis. Results : Treatment of APE increased relative cell viability, prevented a decline of B-cell lymphoma-extra large (Bcl-xL) and cleavage of poly(ADP-ribose) polymerase (PARP) and procaspase-3, and also protected mitochondrial membrane permeability (MMP) against oxidative stress induced by AA+iron. In addition, APE treatment increased phosphorylation of AMP-activated protein kinase (AMPK) exerts a cytoprotective effect. Conclusions : This results demonstrate that APE has an ability to activation of AMPK which protects cells from AA+iron-induced oxidative stress and restores MMP.

• The Korea Journal of Herbology
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• v.26 no.3
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• pp.47-56
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• 2011

Objectives : This study investigated whether the water extract of Spatholobi Caulis (SCE) has the ability to protect hepatocyte against oxidative stress induced by tert-butylhydroperoxide (tBHP) in vitro and $CCl_4$ in vivo. Methods : In vitro, HepG2 cells pre-treated with Spatholobi Caulis water extract (1, 3, 10, $30{\mu}g$/ml) for 12h and further incubated with tBHP ($100{\mu}M$) for the next 12h. Cell viability was assessed by MTT assay. In vivo, rats were orally administrated with the aqueous extract of Spatholobi Caulis (SCE; 50, 100 mg/kg) for 4 days and then, injected with $CCl_4$ 1 mg/kg body weight to induce acute liver damage. Results : Treatment with SCE inhibited cell death induced by tBHP, as evidenced by alterations in the levels of the proteins associated with apoptosis:SCE prevented a decrease in $Bcl_2$, and cleavage of poly(ADP-ribose)polymerase and pro-caspase-3. Moreover, SCE inhibited the ability of tBHP to generate $H_2O_2$ production, thereby restoring GSH content. Moreover, SCE treatments in rats effectively decreased liver injuries induced by a single dose of $CCl_4$, as evidenced by decreases in hepatic degeneration and inflammation as well as plasma alanine aminotransferase and lactate dehydrogenase activities. Consistently, treatments of SCE also protected liver in rats stimulated by $CCl_4$, as indicated by restoration GSH and prevention of MDA in the liver. Conclusions : SCE has the ability 1) to protect hepatocyte against oxidative stress induced by tBHP and 2) to prevent $CCl_4$-inducible acute liver toxicity. Present findings may be informative not only in elucidating the pharmacological mechanism of Spatholobi Caulis, but in determining its potential application for oxidative cellular damage in the liver.

• International Journal of Oral Biology
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• v.39 no.2
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• pp.65-73
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• 2014

Chios Gum Mastic (CGM) is a natural resin extracted from the leaves of Pistacia lentiscus, a plant endemic to the Greek island of Chios. It has been used by traditional healers, and it has antibacterial, antifungal properties, and therapeutic benefits for the skin. The CGM reduces the formation of dental plaque and bacterial growth in oral saliva, and recent studies have demonstrated the role of antioxidant activity of CGM. Although CGM has been widely investigated, its protective effect against oxidative-damage to keratinocytes, as well as the relationship between CGM and autophagy, has not been investigated. The aim of this study was to assess the protective effect of CGM against $H_2O_2$-induced oxidative stress and to evaluate the autophagic features induced by CGM in human keratinocytes. The pretreatment with CGM significantly reduced apoptosis in $H_2O_2$-exposed HaCaT cells. It promoted the degradation of caspase-3, caspase-8, and caspase-9; and it induced the formation of the processed PARP. The treatment with CGM caused an increase in vesicle formation compared to control group. The level of p62 was reduced and the conversion of LC3-I to LC3-II was increased in CGM treated HaCaT cells. Also, the treatment with CGM increased cleavage of ATG5-ATG12 complex. In summary, CGM helps the cells to survive under stressful conditions by preventing apoptosis and enhancing autophagy. Besides, the present investigation provides evidence to support the antioxidant potential of CGM in vitro and opens up a new horizon for future experiments.

• Journal of Microbiology and Biotechnology
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• v.31 no.12
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• pp.1732-1740
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• 2021

Tobacco etch virus protease (TEVp) is a useful tool for removing fusion tags, but wild-type TEVp is less stable under oxidized redox state. In this work, we introduced and combined C19S, C110S and C130S into TEVp variants containing T17S, L56V, N68D, I77V and S135G to improve protein solubility, and S219V to inhibit self-proteolysis. The solubility and cleavage activity of the constructed variants in Escherichia coli strains including BL21(DE3), BL21(DE3)pLys, Rossetta(DE3) and Origami(DE3) under the same induction conditions were analyzed and compared. The desirable soluble amounts, activity, and oxidative stability were identified to be reluctantly favored in the TEVp. Unlike C19S, C110S and C130S hardly impacted on decreasing protein solubility in the BL21(DE3), but they contributed to improved tolerance to the oxidative redox state in vivo and in vitro. After two fusion proteins were cleaved by purified TEVp protein containing double mutations under the oxidized redox state, the refolded disulfide-rich bovine enterokinase catalytic domain or maize peroxidase with enhanced yields were released from the regenerated amorphous cellulose via affinity absorption of the cellulose-binding module as the affinity tag.

• The Korea Journal of Herbology
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• v.27 no.6
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• pp.1-6
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• 2012

Objectives : ROS are involved in a wide spectrum of diseases including chronic inflammation and cancer. S.chinensis Baill, a perennial herb commonly called Chinese lizard's tail or Sam-baek-cho in Korea, is used for the treatment of edema and inflammatory diseases in the Oriental folk medicine. In this study, we investigated the antioxidant activities and anti-inflammatory effects of the two extracts, water(WE) and ethyl acetate(EAE) from S.chinensis Baill. Methods : Anti-oxidant activity was evaluated using Fe2+ chelating and hydroxyl radical scavenging assay. DNA cleavage assay, and western blot and immunostaining for phospho-p65 were performed to evaluate anti-oxidative effect. Anti-inflammatory effect was performed using NO generation assay and western blot in LPS-stimulated RAW264.7 cell. Results : In Fe2+ chelating activity and hydroxyl radical scavenging activity, WE showed more strong scavenging activity for hydroxyl radical than EAE. WE scavenged hydroxyl radical by 12% at 3.2 ${\mu}g/ml$, 21% at 16 ${\mu}g/ml$, 32% at 80 ${\mu}g/ml$, 66% at 400 ${\mu}g/ml$ and 82% at 2000 ${\mu}g/ml$, respectively. In addition, WE showed more strong chelating activity than EAE. WE chelated Fe2+ ion by 1.1% at 3.2 ${\mu}g/ml$, 8.2% at 16 ${\mu}g/ml$, 26.3% at 80 ${\mu}g/ml$, 72% at 400 ${\mu}g/ml$ and 89% at 2000 ${\mu}g/ml$, respectively. Also, WE inhibited oxidative damage via its anti-oxidant activity. In anti-inflammatory effect, EAE inhibited NO production and iNOS expression. In addition EAE suppressed the NF-${\kappa}B$ and MAPK signaling pathway in LPS-stimulated RAW 264.7 cells. Conclusions : Together, these data indicate that S. chinensis Baill, shows anti-oxidant activity and anti-inflammatory effect.