• Toxicological Research
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• v.34 no.3
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• pp.267-279
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• 2018

Neurolathyrism is a neurodegenerative disorder characterized by spastic paraplegia resulting from the excessive consumption of Lathyrus sativus (Grass pea). ${\beta}$-N-Oxalyl-L-${\alpha},{\beta}$-diaminopropionic acid (L-ODAP) is the primary neurotoxic component in this pea. The present study attempted to evaluate the proteome-wide alterations in chick brain 2 hr and 4 hr post L-ODAP treatment. Proteomic analysis of chick brain homogenates revealed several proteins involved in cytoskeletal structure, signaling, cellular metabolism, free radical scavenging, oxidative stress and neurodegenerative disorders were initially up-regulated at 2 hr and later recovered to normal levels by 4 hr. Since L-ODAP mediated neurotoxicity is mainly by excitotoxicity and oxidative stress related dysfunctions, this study further evaluated the role of L-ODAP in apoptosis in vitro using human neuroblastoma cell line, IMR-32. The in vitro studies carried out at $200{\mu}M$ L-ODAP for 4 hr indicate minimal intracellular ROS generation and alteration of mitochondrial membrane potential though not leading to apoptotic cell death. L-ODAP at low concentrations can be explored as a stimulator of various reactive oxygen species (ROS) mediated cell signaling pathways not detrimental to cells. Insights from our study may provide a platform to explore the beneficial side of L-ODAP at lower concentrations. This study is of significance especially in view of the Government of India lifting the ban on cultivation of low toxin Lathyrus varieties and consumption of this lentil.

• Biomolecules & Therapeutics
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• v.19 no.1
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• pp.118-125
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• 2011

Free radical scavenging and antioxidants have attracted attention as a way to prevent the progression of Parkinson's disease (PD). This study was carried out to investigate the effects of n-hexane fraction from Laurus nobilis L. (Lauraceae) leaves (HFL) on dopamine (DA)-induced intracellular reactive oxygen species (ROS) production and apoptosis in human neuroblastoma SH-SY5Y cells. Compared with apomorphine (APO, $IC_{50}=18.1\;{\mu}M$) as a positive control, the HFL $IC_{50}$ value for DA-induced apoptosis was $3.0\;{\mu}g/ml$, and two major compounds from HFL, costunolide and dehydrocostus lactone, were $7.3\;{\mu}M$ and $3.6\;{\mu}M$, respectively. HFL and these major compounds significantly inhibited ROS generation in DA-induced SH-SY5Y cells. A rodent 6-hydroxydopamine (6-OHDA) model of PD was employed to investigate the potential neuroprotective effects of HFL in vivo. 6-OHDA was injected into the substantia nigra of young adult rats and an immunohistochemical analysis was conducted to quantitate the tyrosine hydroxylase (TH)-positive neurons. HFL significantly inhibited 6-OHDA-induced TH-positive cell loss in the substantia nigra and also reduced DA induced $\alpha$-synuclein (SYN) formation in SH-SY5Y cells. These results indicate that HFL may have neuroprotective effects against DA-induced in vitro and in vivo models of PD.

• Biomolecules & Therapeutics
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• v.21 no.1
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• pp.72-78
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• 2013

This work aimed to assess some pharmacological activities of P. leptostachya var. asiatica Hara. The dried roots of P. leptostachya var. asiatica Hara were extracted with 70% ethanol to generate the powdered extract, named PLE. Anti-angiogenic activity was detected using chick chorioallantoic membrane (CAM) assay. In vitro anti-inflammatory activity was evaluated via analyzing nitric oxide (NO) content, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Antioxidant activity was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and reactive oxygen species (ROS) level in the stimulated macrophage cells. Matrix metalloproteinase-9 (MMP-9) and -2 (MMP-2) activities in the culture media were detected using zymography. PLE exhibits an anti-angiogenic activity in the CAM assay, and displays an inhibitory action on the generation of NO in the LPS-stimulated macrophage cells. In the stimulated macrophage cells, it is able to diminish the enhanced ROS level. It can potently scavenge the stable DPPH free radical. It suppresses the induction of iNOS and COX-2 and the enhanced MMP-9 activity in the stimulated macrophage cells. Both monooxygenase and oxidase activities of tyrosinase were strongly inhibited by PLE. Taken together, the dried roots of P. leptostachya var. asiatica Hara possess anti-angiogenic, anti-inflammatory, antioxidant and skin whitening activities, which might partly provide its therapeutic efficacy in traditional medicine.

• Natural Product Sciences
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• v.15 no.1
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• pp.37-43
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• 2009

Oxidative stress is caused by an imbalance between the production of reactive oxygen and an ability of a biological system, to readily detoxify the reactive intermediates or easily repair the resulting damage. It has been suggested that developmental alloxan-induced liver damage is mediated through increases in oxidative stress. The anti-diabetic effect and antioxidant activity of Phaleria macrocarpa (PM) fractions were investigated in alloxan-induced diabetic rats. After two weeks administration of PM, the liver antioxidant enzyme and hyperglycemic state were evaluated. The results showed that oral administration of PM treatments reduced blood glucose levels in diabetic rats by oral administration (P < 0.05). Serum glutamic-oxaloacetic transaminase (sGOT) and serum glutamic-pyruvate-transaminase (sGPT) were also diminished by PM supplementation. The superoxide dismutase (SOD), catalase (CAT) and glutathione-peroxidase (GPx) activities, and glutathione (GSH) level in the alloxan-induced diabetic rats were significantly decreased (P < 0.05) compared to those in the normal rats but were restored by PM treatments. PM fractions also repressed the level of malondialdehyde (MDA) in the liver. Glutathione reductase (GR), glutathione-S-transferase (GST) and $\gamma$-glutamylcysteine synthase (GCS) were also reduced in alloxan-induced diabetic rats. PM fractions could restore the GR and GST activities, but the GCS activity was not affected in rat livers. From the results of the present study, the diabetic effect of the butanol fraction of PM against alloxan-induced diabetic rats was concluded to be mediated either by preventing the decline of hepatic antioxidant status or due to its indirect radical scavenging capacity.

• Natural Product Sciences
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• v.9 no.2
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• pp.68-72
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• 2003

Quercetin and its sugar conjugates are the most abundantly distributed bioflavonoids and represent the largest proportion of flavonols in the plant kingdom. The present study was undertaken to demonstrate the effect of quercetin on the role of reactive oxygen species (ROS) in the development of gastric ulcers in rats. Administration of quercetin in doses of 50, 100 and $200\;mg\;kg^{-1}$ twice daily for 5 days, showed dose dependent significant protection against ethanol (EtOH), aspirin (ASP), cold-restraint stress (CRS) and pylorus ligation (PL) -induced gastric ulcer models and the results were comparable with those elicited by sucralfate. The thiobarbituric acid reactive substances in the stomach mucosa, an index of lipid peroxidation and regulation of plasma corticosterone were significantly increased in CRS-induced gastric ulceration. The queroetin $(100\;mg\;kg^{-1})$ and reduced glutathione effectively inhibited gastric lesions induced by CRS with a significant decrease in the lipid peroxidation and plasma corticosterone. These results indicate that quercetin a bioflavonoid exerts its antiulcer effect in light of free radical scavenging and plasma corticosterone in cold restraint stress ulcers.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.387-397
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• 2003

During the first half of twentieth century, even though the importance of non-calorie essential micronutrients of 13 vitamins and 17 minerals has been known to alleviate nutritional disorder; the primary objective of agriculture and plant breeding programs has been to increase the productivity and seed yields, and macronutrients of proteins, fats, and carbohydrates made up the bulk of foodstuff which were used primarily as an energy source. In the last decade it has been found that non-essential micronutrients encompass a vast group of phytochemicals including antioxidants that are not strictly required in the diet but when present at sufficient levels work as health-promoting chemicals. Nowadays agricultural crops are grown for health rather than for food or fiber, and modifying the nutritional compositions of plant foods has become an urgent health issue. To ensure an adequate intake of essential vitamins and minerals, and to increase the consumption of health-promoting phytochemicals, the researches on plant secondary metabolism have been made. The attempt to improve nutritional quality of crops has been blocked by a lack of basic knowledge of plant metabolism. The advent of genomics era enabled new approaches to make crossing regardless of species, family, or phylum barriers, and the accumulation in our basic knowledge on plant secondary metabolism during the coming decade would be tremendous. As the major staple crops contain insufficient amount of many micronutrients, fortification strategy will be a necessary practice. Elevated intake of specific vitamins, C, E, and $\beta$-carotene, mineral selenium, antioxidants, and phytochemicals significantly reduces the risk of chronic disease such as cancer, cardiovascular disorder, diabetis, and other degenerative disease associated with aging. As the attempt to improve the nutritional quality of crops requires the basic knowledges on plant metabolism, plant biochemistry, human physiology, and food chemistry, strong interdisplinary collaboration among plant biotechnologists, human nutritionists, and food scientists will be needed. Inhibition of cancer, cardiovascular disease, and other degenerative disorder may be the biggest goal facing nutritional plant breeders. But the assumption that simply increasing dietary level of any compound will necessarily improve human health is a dangerous idea because many plant secondary products and dietary contaminants have paradoxical (hermetic) effects. Before biotechnical manipulation is undertaken to elevate or reduce any individual constituent of crops, the contribution of the micronutrient to human health must first be investigated.

• Journal of Microbiology and Biotechnology
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• v.24 no.4
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• pp.483-488
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• 2014

In order to improve the expression of heat-resistant xylanase XYNB from Aspergillus niger SCTCC 400264, XynB has been cloned into Pichia pastoris secretary vector pPIC9K. The XynB production of recombinant P. pastoris was four times that of E. coli, and the $V_{max}$ and specific activity of XynB reached $2,547.7{\mu}mol/mg$ and 4,757 U/mg, respectively. XynB still had 74% residual enzyme activity after 30 min of heat treatment at $80^{\circ}C$. From the van der Waals force analysis of XYNB (ACN89393 and AAS67299), there is one more oxygen radical in AAS67299 in their catalytic site, indicating that the local cavity is much more free, and it is more optimal for substrate binding, affinity reaction, and proton transfer, etc, and eventually increasing enzyme activity. The H-bonds analysis of XYNB indicated that there are two more H-bonds in the 33rd Ser of XYNB (AAS67299) than in the 33rd Ala(ACN89393 ), and two H-bonds between Ser70 and Asp67.

• Journal of Environmental Health Sciences
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• v.27 no.2
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• pp.10-16
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• 2001

To evaluate an effect of circadian variation on the xylene metabolizing enzyme activities, 50% m-xylene in olive oil(0.25 $m\ell$/100g body weight) was intraperitoneally administered to the rats every other day for 6 days both in the night; 24:00 and the day; 12:00. Then animals were sacrigiced at 8hr after last injection of m-xylene. Hepatic microsomal cytochrome p450 contents were more increased both in control and xylene treated rats of night phase than those of day phase. But the activity of hepatic alcohol dehydrogenase(ADH) in control of night phase showed the similar value with that in those of day phase and xylene treated rats of day phase showed an increasing tendency of hepatic ADH activity as those of night phase showing similar activity. Furthermore, control rats of night phase than those of day phase. And by xylene treatment, enzyme activities of rats of day phase were higher tendency in rats of control but those of night phase were somewhat inhibited. Besides, xylene-treated animals of night phase showed increasing tendency of urinary methylhippuric acid concentration compared with those of day phase. On the other hand, liver weight per body weight(%), hepatic lipid peroxide content and serum xanthine oxidase activity were higher in night phase. And the activities of hepatic oxygen free radical metabolizing enzymes such as xanthine oxidase, gluthathione S-transferase, and xylene-treated rats of night phase than those of day phase. In conclusion, it can be hypothesized on the basis of the results that the accumulation rate of m-xylene intermediate metabolite, i.e. m-methylbenzaldehyde in liver tissus may be higher in night phase than in day phase and it may be responsible for higher liver toxicity in bight phase than in day phase.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.23-31
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• 2015

Catalpol, an iridoid glucoside, isolated from the root of Rehmannia glutinosa Libosch. It possesses a broad range of biological and pharmacological activity including anti-tumor, anti-inflammation and anti-oxidant by acting as a free radical scavenger. Therefore, in this study, the effects of catalpol on blastocyst development, expression levels of reactive oxygen species (ROS) and apoptotic index were investigated in porcine embryos. After in vitro maturation and fertilization, porcine embryos were cultured for 6 days in porcine zygote medium 3 (PZM-3) supplemented with catalpol (0, 100, 200 and $400{\mu}M$, respectively). Blastocyst development not significantly improved in the catalpol treated group when compared with control group. Otherwise, the intracelluar levels of ROS were decreased and the numbers of apoptotic nuclei were reduced in the catalpol ($100{\mu}M$) treated porcine blastocysts (P<0.05). On the other hand, blastocyst development was significantly improved in the catalpol ($100{\mu}M$) treated group when compared with the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Otherwise, the intracellular levels of ROS in catalpol ($100{\mu}M$) treated group were significantly decreased in the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Furthermore, the total cell numbers of blastocysts were significantly increased (P<0.05) in the catalpol ($100{\mu}M$) treated group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress, whereas numbers of apoptoic nuclei were significantly reduced (P<0.05). In conclusion, our results indicate that treatment of catalpol may have important implications for improving developmental competence and preimplantation quality of porcine embryos through its anti-oxidant and anti-apoptotic effect.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.4
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• pp.1031-1036
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• 2003

To certify the protective effect of herbal medicine against oxygen free radical-induced myocardiotoxicity, cytotoxicity was measured using TBARS assay and Beating rate in the presence of Tongryeong-san(TRS) extracts or single constituents of this prescription. Myocardial toxicity was evaluated in neonatal rat myocardiocytes in cultures. In the present study, xanthine oxidase/hypoxanthine (XO/HX) resulted in a increase in lipid peroxidation and decreases in beating rate in cultured myocardial cells. In the effect of TRS extract, it showed the prevention from the XO/HX-induced cardiotoxicity by the increases of beating rate as well as the decrease of lipid peroxidation, In the protective effect of Faeces Trogopterori(FT), Pollen Typhae(PT), Caulis Akebiae(CA) and Radix Paeoniae Rubra(PRR), all the extracts were significantly effective in the protection of XO/HX-induced cardiotoxocity in cultured myocardial cells by the increase of beating rate as well as th decrease of lipid peroxidation. From these results, they show that XO/HX is cardiotoxic in cultured myocardial cells derived from neonatal rat, and it suggests that TRS, FT, PT, CA and PRR extracts are positively effective in the blocking in XO/HX-induced cardiotoxicity.